Ginsenosides are the abundant secondary metabolites in American ginseng (Panax quinquefolium), it could be released into soil through root exudation and decomposition during plant growth. This study determined ginsenoside contents in American ginseng cultivated soil by HPLC. Three ginsenosides, Rb1, Rb2 and Rd, were detected in the rhizosphere soil of 3-4 years old American ginseng cultivated in Huairou District, Beijing, and their contents were 0.80-3.19 mg x kg(-1). Correspondingly, the contents of the three ginsenosides in soil solution were 4-16 mg x L(-1) at field water-holding capacity of 20%. According to the field soil test data, we designed the concentration of ginsenosides for bioassays (0.2-125 mg x L(-1) in solution or 0.2-125 mg x kg(-1) in soil). The results showed that radicle lengths of American ginseng were reduced by 6%-23% in solution containing 0.2-125 mg x L(-1) ginsenoside extract, and a significant difference was observed at concentration of 125 mg x L(-1) (P < 0.05). The shoot lengths of American ginseng were not significantly inhibited by 0.2-125 mg x L(-1) ginsenosides extractions. After 20 days of growth in nutrient solution amended with 25 mg x L(-1) ginsenosides extraction, plant height of 3-year-old American ginseng seedling was decreased by 28% compared to the control, and the biomass of aerial parts was also reduced by 50% (P < 0.05). However, the growth of newly-grown fibrous root was not significantly inhibited. Comparatively, when American ginseng embryos were cultivated into sterile or non-sterile soil, neither radicle lengths nor shoot lengths were significantly affected by 0.2-125 mg x kg(-1) ginsenoside extracts. In conclusion, ginsenosides showed autotoxic effect on growth of American ginseng radicle and adult seedling, however, this effect was weakened in field soil.
Chromatography, High Pressure Liquid ; Culture Media ; chemistry ; metabolism ; Ginsenosides ; analysis ; metabolism ; toxicity ; Panax ; chemistry ; drug effects ; growth & development ; metabolism ; Plant Roots ; chemistry ; drug effects ; growth & development ; metabolism ; Soil ; chemistry

Network pharmacology is an emerging discipline based on the Disease-Gene-Drug multilevel network. And it has been used to forecast the drug targets and improve the efficiency of drug discovery. Its research ideas are similar to the overall efficacy of traditional Chinese medicine (TCM), which attracts more and more medical re-searchers to look for the joint point of TCM and network pharmacology. A series of approaches on disease-related genes, predicting the information of target and active ingredients of TCM emerge. In this paper, the network pharma-cology research tools, databases and their applications were summarized and introduced. This paper also proposed scientific strategies to separate active ingredients of TCM using network pharmacology, so as to improve the efficiency and speed of finding active ingredients of TCM.

Objective To observe the changes of coagulation function in patients with acute lowerlimb deep venous thrombosis(DVT)and evaluate the risk factors for DVT. Methods Plasma APTT.PT,TT,D-dimer and fibrinogen(Fbg)were detected by an automated coagulation analyzer in 62 acute lower-limb DVT patients and 70 controls:Retrospective studies on the clinical data of all patients were done by binary logistic regression analysis.Results (1)In DVT group,plasma APTT,PT and TT,the levels of D-dimer and fibrinogen.and D-dimer/fibrinogen ratio(D/F ratio)were higher when compared with control group(au P＜0.01);(2)There were positive correlations between D-dimer and fibrinogen both in DVT and control groups(r=0.475,P＜0.01;r=0.564,P＜0.01,respectively);(3)Logistic analysis indicated that acute lower-limb DVT was associated with the presence of hypertension and increased plasma level of fibrinogen(OR=24.99,P＜0.01: OR=4.346.P＜0.01,respectively).Conclusions Hypertension and elevated plasma level of fibrinogen are independent risk factors for acute lower-limb DVT.

AIMTo explore the effects of sinomenine(Sin) on intracellular free calcium ([Ca2+]i) and the activity of PKC (protein kinase C) of the cultured aortic vascular smooth muscle cells (VSMC) during ischemia and hypoxia.

METHODSThe effect of Sin on changes in [Ca2+]i were determined in cultured rabbit VSMC after exposure to high K+, norepinephrine (NE) and caffeine (Caf). Fluorescent Ca2+ -indicater fura-2/AM was used. The effects of Sin were compared with that of verapamil (Ver). The hypoxia model was made, then the activity of PKC was measured by y scintillation counting instrument.

RESULTSSin (10 x 10(-6) mol x L(-1), 3 x 10(-5) mol x L(-1) 10(-4) mol x L(-1)) inhibited the elevation of [Ca2+]i induced by high K+ -depolarization in a concentration dependent manner. In addition, Sin inhibited the elevation of [Ca2+]i induced by NE in the presence of extracellular Ca2+. In the absence of extracellular Ca2+, Sin (3 x 10(-5) mol.L(-1)) also had no blocking effect on the NE-induced [Ca2+]i increase. It was found that the activity of PKC treated with Sin in VSMC cytoplasm and cell membrane in normal condition increased, the activity of PKC in cytoplasm in ischemia and hypoxia situation increased, but the activity of PKC in cell membrane decreased. When VSMC was treated with Sin, the activity of PKC in cytoplasm decreased and that of cell membrane increased.

CONCLUSIONThe results suggest that Sin might decrease the[Ca2+] i of VSMC by blocking both VDC and ROC, could regulate the PKC activities induced by ischemia and hypoxia.

Animals ; Aorta ; cytology ; drug effects ; Calcium ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Cytoplasm ; metabolism ; Morphinans ; pharmacology ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; physiology ; Protein Kinase C ; metabolism ; Rabbits

Objective:To analyze the result of smooth pursuit test(SPT) in unilateral vestibular peripheral vertigo and investigate its influencing factors.Method:Smooth pursuit test (SPT) and spontaneous nystagmus (SN)were examined in one hundred and eighty-five patients with unilateral peripheral vertigo(case group) and 51 normal persons(control group) by Video-Nystagmography (Synapsys, France), and the gain of SPT and SN were selected as the observation parameters in order to analyze the waveform and gain of SPT and the relativity between SN and the gain of SPT.Result:Of the 185 patients, 105(56.8%),72(38.9%) and 8(4.3%) cases producedⅠ,Ⅱ and Ⅲ waveforms respectively. Of these patients, 58(31.4%) demonstrated SN and none had Ⅳ waveform. While of 51 normal persons, 38(74.5%), 13(25.5%) persons producedⅠand Ⅱwaveforms repectively and there were no Ⅲ, Ⅳ waveforms or SN. There was statistical significance between the stong and weak gain of SPT in these two groups. Weak gain was significantly different between two groups. The stong and weak gain of SPT in case group were 0.86±0.06,0.80±0.06; 0.78±0.09, 0.65±0.1; 0.68±0.13, 0.45±0.12. The relativity between SN and the gain of SPT was positive when they had same direction(r_s=-0.63,P＜0.05)and negative when opposite (r_s=0.34,P＜0.05).Conclusion:Ⅰ,Ⅱ,Ⅲ three waveforms of SPT could appear in unilateral vestibular peripheral vertigo and the corresponding gains are gradually decreasing.SN is the influencing factor of SPT.

AIMTo survey the uptake behavior and subcellular distribution of antisense oligodeoxynucleotide polymethacrylate submicroparticles (AS-ODN-SMP) and infer its mechanism in MGC cell lines.

METHODSMGC cells were incubated at certain concentration of AS-ODN-SMP or AS-ODN for 8 h at 4 degrees C or 37 degrees C. Then the fluorescence oligodeoxynucleotide- labeled cells were counted by flow cytometer and the intracellular fluorescence intensity was determined after incubated with chloroquine for 2 h.

RESULTSCellular uptake of oligodeoxynucleotides was significantly increased following application of AS-ODN-SMP and total intracellular fluorescence intensity was enhanced by 683 folds with the vehicle concentration of 20 microg x mL(-1). AS-ODN-SMP entranced to cells profoundly with temperature-dependent manner. Rare cells took on fluorescence when incubated at 4 degrees C, while 37 degrees C they were significantly increased. But the intracellular fluorescence intensity appeared same level in present or absent of chloroquine.

CONCLUSIONWith the help of polyacrylate submicroparticles, oligonucleotides efficiently entranced the cells via endocytosis and could successfully escape the degradation in lysosome.

Animals ; Cell Line ; Drug Carriers ; Drug Delivery Systems ; Endocytosis ; drug effects ; Giant Cells ; cytology ; Lysosomes ; metabolism ; Nanoparticles ; Oligodeoxyribonucleotides, Antisense ; administration & dosage ; pharmacokinetics ; Particle Size ; Polymethacrylic Acids ; chemistry ; pharmacology ; Temperature

OBJECTIVE: To observe cardiac ultrastructure and the expression of heat shock protein 70 (HSP70) and hypoxia inducible factor-lα (HIF-lα) in electric shock death rats and to explore the application of these indexes as the basis of medical identification in electric shock death. METHODS: Seventy-two SD rats were randomly divided into electric shock death group, postmortem electric shock group and the control group. The changes of myocardial ultrastructure were observed by transmission electron microscope, and the expressions of myocardial HSP70 and HIF-1α were observed by immunohistochemical technology. RESULTS: Myocardial myofibril fracture, mitochondrial cristae and membrane dissolution, and disordered arrangement of Z lines and M lines were observed in electric shock rats. HSP70 and HIF-lα were strong positive expressions in the electric shock death group, significantly compared with the control and postmortem electric shock groups (P < 0.05). CONCLUSION The expressions of HSP70 and HIF-lα were obviously increased in electric shock death group, which may be used as the diagnostic indicator of electric shock death.
Animals ; Death ; HSP70 Heat-Shock Proteins/metabolism* ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Myocardium/pathology* ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of nerve growth factor (NGF) on lipopolysaccharide (LPS) injury and activation of nuclear factor-kappa B in PC12 cells.

METHODSIn order to set injury models, the PC12 cells were incubated in different concentration of LPS. Cells were cultured in the culture and were reduced by LPS, and then cells were treated by NGF of various concentrations. The cell viability was determined by methyl thiazolyl tetrazolium (MTT) assay, cellular morphology was observed under inverted microscope and fluorescence microscope, and the content of NF-kappaB was assessed by RT-PCR.

RESULTS(1) The viability of PC12 cell was decreased with concentration of LPS increasing. (2) The cellular morphology change showed that NGF had an ability to reduce LPS injury. (3) The result of RT-PCR showed that the content of NF-kappaB in LPS injury was more than the normal and treated cell, and the treated one was close to the normal one.

CONCLUSIONThe reports about NGF in brain cells repair after inflammatory are very small. And our study is about that NGF can protect the PC12 cell from LPS injury, and this mechanism possible bears on the activation of NF-kappaB.

Animals ; Lipopolysaccharides ; toxicity ; NF-kappa B ; metabolism ; Nerve Growth Factor ; pharmacology ; Neuroprotective Agents ; pharmacology ; PC12 Cells ; Rats

OBJECTIVETo prepare OANO-1 microspheres and test their release in vitro.

METHODOANO-1 microspheres were made by W/O/W-liquid drying process. The surface morphology of the microspheres was observed by SEM. The mean diameter and the size distribution of microspheres, the drug loading and the incorporation efficiency were examined. The release of OANO-1 microspheres in vitro was examined by small cup method. The accumulated release percent of OANO-1 microspheres was examined.

RESULTThe OANO-1 microspheres were regular in their morphology. The average particle size was 8.59 microm with over 90% of the microspheres being in the range of 1-12 microm. The drug loading and the incorporation efficiency were 48.39% and 19.32% respectively. The accumulated release percent of OANO-1 microspheres was 78.4% after 108 h. The release half-life t1/2 was 40.8 h and Higuchi equation was Y = 0.1326 X - 0.4782, r = 0.9951.

CONCLUSIONThe preparation of OANO-1 microspheres was well. The release in vitro of OANO-1 microspheres showed significant sustained release.

Angelica sinensis ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; Drug Combinations ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Epimedium ; chemistry ; Ficusin ; analysis ; Furocoumarins ; analysis ; Microspheres ; Particle Size ; Plants, Medicinal ; chemistry ; Psoralea ; chemistry

AIMTo prepare lung targeting azithromycin cationic liposomes and to observe its tissue distribution in mice.

METHODSThe azithromycin cationic liposomes were prepared by thin film method with freeze-thawing steps. HPLC method was established and validated for the determination of azithromycin in tissues of mice.

RESULTSThe particle size of the liposomes was 6.582 microm with zeta potential of +19.5 mV. The entrapment efficiency was more than 75%. The liposomes was stable in 6 months stored at 4 degrees C. The release in vitro was characterized by Higuchi equation. Azithromycin liposomes and free azithromycin solution were injected intravenously at a dose of 80 mg x kg(-1) to mice. Compared with solution, liposomes were characterized by slower clearance, increased half-life and the AUC increased by 7.4 fold in lung.

CONCLUSIONThin film method with freeze-thawing steps could increase the entrapment efficiency and increase the particle size of azithromycin liposomes. After modification of lipid membrane with stearylamine, the cationic liposomes were prepared. The azithromycin concentration and AUC increased in lung after iv administration to mice of the cationic liposomes. This offered a good information for preparing liposomes targeting on the lung.

Animals ; Anti-Bacterial Agents ; administration & dosage ; chemistry ; pharmacokinetics ; Area Under Curve ; Azithromycin ; administration & dosage ; chemistry ; pharmacokinetics ; Cations ; Drug Carriers ; Drug Compounding ; methods ; Drug Delivery Systems ; Female ; Liposomes ; Lung ; metabolism ; Male ; Mice ; Particle Size ; Tissue Distribution