OBJECTIVEEffects of ginsenoside Rb1, Rg1 and Re on neurotrophic factor signal transduction pathway using liposome-mediated transfection of eukaryotic cells approach.

METHODThe injury model was established by treating SH-SY5Y cells with 0.6 mmol x L(-1) of corticosterone (CORT) by 24 h. SH-SY5Y cell were pretreated with CORT for 30 min followed by co-treated with 120,60 and 20 micromol x L(-1) of Rb1, 120, 80 and 40 micromol x L(-1) of Rg1 and 120, 80 and 40 micromol x L(-1) of Re for 24 h. Cells viability was determined by Cell Counting Kit (CCK) assay. CREB expressing Luciferase reporter gene was constructed and transfected with plasmid containing hRaf, hcAMP, hAkt, hCaMK gene into human embryonic kidney (HEK293) cells using liposornal transfection reagent lipofection 2000. The expression of CREB before and after it addion of Rb1, Rg1 and Re was examined by Luc assay system and Western blotting.

RESULTCompared with normal control group, CORT significantly decreased the viability of SH-SY5Y cells to 67.21% (P < 0.01). CCK results show that Rb1 (60 micromol x L(-1)), Rg1 (80 micromol x L(-1)) and Re (80 micromol x L(-1)) on SH-SY5Y cells have significant protective effect (P < 0.01). Lucassay and Western blotting results show that the gene and protein levels of CREB increased significantly through the pathway of Raf and Akt with Rb1 and Rg1 (P < 0.01), Re can increase significantly the gene and protein levels of CREB through the pathway of Raf and CaMK II.

CONCLUSIONRb1, Rg1 and Re protects SH-SY5Y cells from CORT-induced damage and the neuroprotective mechanism may be associated with the Raf-CREB, Akt-CREB and CaMK II -CREB pathways.

Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; genetics ; metabolism ; Cell Line ; Cell Survival ; drug effects ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Genes, Reporter ; Ginsenosides ; pharmacology ; Humans ; Panax ; chemistry ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Signal Transduction ; drug effects ; raf Kinases ; genetics ; metabolism

Objective To explore the effects of chronic fluorosis on neurons in the cerebral cortex of rats,and to provide some morphological evidence of damage in the central nervous system induced by chronic fluorosis.Methods Male Wistar rats 40 days after birth were fed with high fluoride contented water(100 mg/L)for inducing chronic fluorosis.Immunocytochemistry and in situ hybridization were used to detect c-fos and Caspase 8 at cerebral cortical neurons respectively.Results c-fos positive cells rate and gray scale in the cerebral cortex of chronic fluorosis were 35.8%and 0.2756±0.0241,respectively,and that of control group were 32.1%and 0.2774±0.0331with statistical difference(χ2=0.305,t=0.826,P＞0.05).Caspase 8 positive cells rates of fluorosis group and control group were 18.7%and 14.1%,respectively,the difference being statistically significant(χ2=0.419,P＞0.05).The gray scale of fluorosis group and control group were 0.3874±0.0329 and 0.3884±0.0323,respectively,the difference being statistically significant(t=0.641,P＞0.05).Conclusion Chronic fluorosis had no significant influence on apoptosis of cerebral cortical neurons.

Objective: To investigate the changes of energy metabolism before and after ginger-processing of Coptidis Rhizoma, and elucidate the relationship between the changes of medicinal properties and the energy metabolism. Methods: Crude Coptidis Rhizoma and ginger-processed Coptidis Rhizoma decoctions were prepared and administered to rats once a day for 7 d by gavage. Blood and liver were used to detect the content and activity of key enzymes in three pathways of energy metabolism. The composition analysis analyzed the detection results and screened out the key enzymes for the main changes in energy metabolism before and after processing. Results: Compared with the control group, the activities of HK, PFK, PK and mitochondrial respiratory chain complex I, II, III and IV in crude Coptidis Rhizoma group were decreased significantly, and the content of CS, ICD and α-KGDH was decreased significantly.The activities of mitochondrial respiratory chain complexes II, III and IV and the activities of PFK and PK in the ginger-processed Coptidis Rhizoma group were significantly decreased. HK and mitochondrial respiratory chain complex I was decreased to some extent, but there was no significant difference. The content of α-KGDH was decreased significantly, and the content of CS and ICD was decreased in a certain degree. Compared with the crude Coptidis Rhizoma group, the activities of HK, PFK, PK and mitochondrial respiratory chain complex I, II, and III in the ginger-processed Coptidis Rhizoma group were increased significantly, and the content of CS, ICD and α-KGDH was increased significantly. There was no significant change in the mitochondrial respiratory chain complex IV. Two principal components were extracted by principal component analysis. Among them, complex II and complex III had the maximum contribution rate of principal component 2 and principal component 1 of 0.916 and 0.873, respectively. Conclusion: The weakening of cold in nature of Coptis chinensis after ginger-processed treatment may be due to the inhibition of the inhibitory effect of C. chinensis on mitochondria respiratory chain complex II and complex III in the oxidative phosphorylation pathway.

Five compounds (tenuifoliside C, tenuifoliside D, telephiose A, telephiose C and polygalaxanthone III) from polygala tenuifolia wild were incubated together with CYP probe substrate in human liver microsomes to investigate the inhibitory effect towards CYP450 enzyme. Phenacetin (CYP1A2), coumarin (CYP2A6), paclitaxel (CYP2C8), diclofenac (CYP2C9), S-mepheriytoin (CYP2C19), dextromethorphan (CYP2D6), chlorzoxazone (CYP2E1), midazolam (CYP3A) were selected as the isoforfn specific substrate. And the formation of paracetamol, 7-hydroxycoumarin, 6alpha-hydroxy paclitaxel, 4'-hydroxydiclofenac, dextrorphan, 6-hydroxychlorzoxazone, 1'-hydroxymidazolam, 4'-hydroxymephenytoin were detected respectively to measure the effect towards CYP450 by high-pressure liquid chromatography (HPLC). The result shows that five compounds from polygala tenuifolia willd significantly inhibit chlorzoxazone 6-hydroxylation catalyzed by CYP2E1, while showed no effect towards CYP1A2, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A. And IC50 value was 38.73, 54.14, 61.77, 62.22, 50.56 micromol x L(-1), respectively.
Cytochrome P-450 Enzyme System ; metabolism ; Esters ; pharmacology ; Glycosides ; pharmacology ; Humans ; Microsomes, Liver ; drug effects ; enzymology ; Oligosaccharides ; pharmacology ; Polygala ; chemistry ; Xanthones ; pharmacology

Objective Using Intelligence Scale of Mini Mental State Estimated (MMSE) as the gold standard to determine the relevance of International HIV-associated Dementia Scale (IHDS)in minority ethnic areas in Guangxi populations with different cultural values.Corresponding boundary value related to the authenticity and reliability on IHDS were also evaluated.Methods 200 patients with HIV infection were randomly selected from the minority ethnic groups in Guangxi.For each infected person,MMSE and IHDS blind scale were tested at the same period.Using the results from MMSE scale test as the gold standard,ROC curve and IHDS scale in Guangxi minority populations with different education levels which related to the diagnosis of dementia-HIV values were determined.The value of a specific sector under the IHDS sensitivity,specificity,and internal consistency coefficients was also evaluated.Results When considering the infected person did not differ on their educational level,the IHDS scale diagnostic cutoff appeared as 8.25,while 1HDS sensitivity as 0.925,specificity as 0.731 and Kappa as 0.477 (P＜0.001).When considering the extent of cultural differences did influence the prevalence of infection,the different education groups showed different IHDS diagnostic cutoff values.People with high school,secondary school or higher education levels,the IHDS diagnosis appeared to be 8.25,when sensitivity was 0.917,specificity was 0.895 and Kappa was 0.722 (P＜0.001).People with only primary education level,the IHDS appeared to be 7.25.When sensitivity was 0.875,specificity was 0.661 and Kappa was 0.372 (P＜0.001).Conclusion The IHDS diagnostic sector in Guangxi minority groups was lower than the internationally recommended level of diagnostic cutoff value (IHDS≤ 10 points).When using IHDS to perform the HIV related dementia screening program,in the minority areas of Guangxi,culture context,the degree and difference of HIV infection should be considered,especially in using IHDS diagnostic cutoff values.

italic>Alangium chinense is a commonly used medicinal plant of Alangiaceae Alangium, one of the characteristic Miao medicines in Guizhou. The genus is strongly differentiated and has complex morphological variation, with significant differences in active ingredients and potency among herbs. In this paper, we sequenced the chloroplast genomes of Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib using Illumina high-throughput sequencing technology. The genomes of Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib chloroplasts were sequenced, their assembly annotation and structural characterization were completed, and the genomes of the congeners Alangium chinense and Alangium alpinum were downloaded from NCBI for comparative analysis. Finally, the phylogenetic tree was constructed by selecting published species with close affinity to Alangium chinense family from NCBI. The results were as follows: Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib chloroplast genomes were 156 670, 156 672 and 156 871 bp in length, with a typical four-segment structure, all containing one long single copy region (LSC), one short single copy region (SSC) and two inverted repeat regions (IRa and IRb). All of them were annotated to 133 genes, including 88 protein-coding genes, 37 tRNA genes and 8 rRNA genes. Three types of long repeat sequences and SSR loci, forward repeats, palindrome repeats and reverse repeats, were found in all chloroplast genomes. Comparative genomic analysis showed that there was diversity in the boundary transition regions of the five species, no rearrangements or inversions were found in the chloroplast genome, and there were significant differences in the coding regions of ndhA, ycf1, rpl16, ycf2, and petD genes, which provided new loci for molecular identification of Patagonia. In the phylogenetic analysis, Alangium kurzii var. kurzii clustered as a single species, and Alangium chinense subsp. Pauciflorum and Alangium chinense subsp. Strigosum clustered as a single species, with a support rate of 93 and close affinity, indicating that the phylogenetic tree can be used for the species identification. This paper can provide a basis for the taxonomic identification of Alangium, ensure the safety of clinical use of anise herbs and regulate the market of Alangium chinense herbs.

To establish an HPLC-MS/MS method for the analysis of quercetin, kaempferid and isorhamnetin in rats plasma and study its pharmamacokinetics after an intragastrical administration of Hippophae rhamnoides extracts. Five healthy male Sprague-Dawley (SD) rats were given single doses of H. rhamnoides extracts (quercetin 26.35 mg x kg(-1), kaempferid 4.040 mg x kg(-1), isorhamnetin 31.37 mg x kg(-1)), and then their orbital sinus blood samples were collected at different time points. The drug plasma concentration of the three flavonoids was determined by HPLC-MS/MS method. After that, the main pharmacokinetics parameters were calculated by using Kinetica 5. 0. 11 software. The methodological test showed that the linear concentration ranges of quercetin, kaempferid and isorhamnetin were 7.500-600.0 μg x L(-1) (R2 = 0.998 5), 1.000-80.00 μg x L(-1) (R2 = 0.998 5 ) and 10.00-800.0 μg x L(-1) (R2 = 0.998 0), respectively. The inner and inter-days precisions were both less than 14.0%. The plasma samples showed a good stability and consistency with the requirement of biological sample analysis after the samples were frozen once and placed at - 20 degrees C for 15 d and room temperature for 6 h and the treated analytes were placed at -20 degrees C for 24 h. For quercetin, the pharmacokinetic parameter t(½β), AUC(0-∞), MRT(0.∞), C.(max) and T(max) were (113.3 ± 19.37) min, (12 542.14 ± 3 504.05) μg x h x L(-1), (119.6 ± 13.29) h, (164.6 ± 27.33) μg x L(-1) and (5.199 ± 0.840 3) h, respectively. For kaempferid, the pharmacokinetic parameters t(½β), AUC(0-t), MRT(0-∞), C(max) and T(max) were (79.85 ± 17.15) min, (934.51 ± 94.59) μg x h x L(-1), (81.50 ± 13.75) h, (80.15 ± 14.24) μg x L(-1) and (3.827 ± 0.902 7) h, respectively. For isorhamnetin, the pharmacokinetic parameters t1,2,, AUC(0-t), MRT(0-∞), C(max) and T(max) were (118.3 ± 20.73) min, (26 067.77 ± 4 124.60) μg x h x L(-1), (129.0 ± 16.30) h, (269.6 ± 29.32) μg x L(-1) and (6.513 ± 1.450) h, respectively. The HPLC-MS/MS analysis method established in this study was proved to be sensitive and accurate and could be applied in the pharmacokinetic study of quercetin, kaempferid and isorhamnetin in rat plasma.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Hippophae ; chemistry ; Kaempferols ; blood ; pharmacokinetics ; Male ; Quercetin ; analogs & derivatives ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

Objective To preliminarily investigate the clinical features and efficacy of surgical treatment of moyamoya syndrome in patients w ith hyperthyroidism. Methods From December 2002 to April 2013, 41 patients w ith moyamoya syndrome based on the disease of hyperthyroidism admitted to the Department of Neurosurgery, the 307th Hospital of PLA w ere analyzed retrospectively. The clinical data w ere colected, including sex, age of onset, initial symptoms, progress symptoms, imaging features, Suzuki staging, and surgical efficacy, and they w ere compared w ith the clinical data of the patients w ith moyamoya disease treated at the same time. Results The ratio of male to female w as about 1∶4 in moyamoya syndrome patients w ith hyperthyroidism. The peak age of onset w as 25 to 34 years old. Compared w ith the patients w ith moyamoya disease at the same period, the proportion of patients w ith cerebral infarction as initial symptom w as higher in the moyamoya syndrome group ( 39.0% vs.24.2%; χ2 = 4.796, P =0.029), more patients had symptomatic progression (46.3% vs.25.4%; χ2 =9.207, P =0.002), and the proportion of of patients w ith cerebral hemorrhage as initial symptom w as low er ( 2.4% vs.14.6%; χ2 = 4.829, P =0.028). Thirty-seven patients w ho received encephalo -duro-arterio-synangiosis (EDAS) w ere folow ed up for 43 ± 19 months. The results show ed that the clinical symptoms of 31 patients had different degrees of improvement. Conclusions Moyamoya syndrome of hyperthyroidism is more common in w omen. The risk of cerebral infarction is higher and more prone to have disease progression. The efficacy of EDAS is better for controling the progression of moyamoya syndrome in patients w ith hyperthyroidism.

OBJECTIVETo compare the effect of reinforcing Shen method (RSM) and activating blood method (ABM) in treating osteoarthritis (OA) at the molecular level.

METHODSThe physical and chemical characteristics of components from respective recipes of RSM and ABM, and network features of component-target interaction network were analyzed by computer simulation methods including chemical space, molecular docking, and biological network, etc.

RESULTSThe chemical components of RSM and ABM were scarcely scattered with larger overlapping. Among established networks, the distribution of network features was partially similar in RSM and ABM. The average target number correlated with each component was 1.86 in RSM and 2.11 in ABM respectively. Each average target number was respectively correlated with 4.46 compounds and 3.93 compounds, reflecting multi-component and multi-target actions.

CONCLUSIONComputer simulation could intuitively trace out similarities and differences of two different methods and their interaction with targets, which revealed that the compatibility of RSM and ABM could have broader protein targets and potential synergism at the molecular level.

Computer Simulation ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Osteoarthritis ; drug therapy ; Phytotherapy ; methods

The effect of plant growth regulator forchlorfenuron (CPPU) 1 x 10(-6), 0.67 x 10(-6), 0.5 x 10(-6) on fruit morphology and effective components lignans was studied. Those morphologies were the combination of four basic morphological changes. The result showed, diametre were increased and longitudinal diametre of fruits were inhibited by foliage fertilizers including CPPU. At the same time, 1 000-grain weight and yield showed the varying degrees increase under CPPU. The order of the degree was 0.5 x 10(-6) > 1 x 10(-6) > 0.67 x 10(-6). Six lignans content of Schisandra chinensis of different harvest time and different CPPU processing groups were determined, the results showed that lignans accumulation occurred mainly in periods of premature the half mature fruiting stages. Under the 0.67 x 10(-6) CPPU treatment, schisandrol B, schisandrin B, schisandrin C content of S. chinensis showed different increase.
Chromatography, High Pressure Liquid ; Cyclooctanes ; analysis ; metabolism ; Dioxoles ; analysis ; metabolism ; Dose-Response Relationship, Drug ; Fruit ; drug effects ; growth & development ; metabolism ; Lignans ; analysis ; metabolism ; Phenylurea Compounds ; pharmacology ; Polycyclic Compounds ; analysis ; metabolism ; Pyridines ; pharmacology