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Objective: To investigate the Daodi habitat of Panacis Majoris Rhizoma by analyzing the characteristics of inorganic elements in Panacis Majoris Rhizoma from different habitats., Methods: The contents of inorganic elements in Panacis Majoris Rhizoma from different habitats were determined by ICP-AES. The characteristics of inorganic elements in Panacis Majoris Rhizoma were analyzed by correlation analysis, principal component analysis and cluster analysis., Results: It was showed that there was a correlation between the contents of inorganic elements and the medicine quality of Panacis Majoris Rhizoma; Fe, Cr, Al, Mg, Cd, Ca and Zn were principal components of Panacis Majoris Rhizoma; and the contents of inorganic elements in Panacis Majoris Rhizoma existed regional differences., Conclusion: The contents of inorganic elements Ca, Fe and Zn,especially the content of the essential trace elements Fe and Zn, can be used as one of the key reference for medicinal quality evaluation of Panacis Majoris Rhizoma; as well, Shaanxi Province is probably the Daodi habitat of Panacis Majoris Rhizoma.

The dysfunction of vascular system is one of the main causes of orthostatic intolerance induced by microgravity. Vascular endothelial cell is a single layer on the inner wall of the blood vessel and is the important component of the blood vessel wall. Vascular endothelial cell plays a pivotal role in the regulation of vascular functions, such as serving as a permeability barrier, regulating vasoconstriction and vasodilatation. Recent studies have demonstrated that microgravity may have different effects on vascular sys- tem and vascular endothelial cells in different parts of the body, such as increasing vasoconstrictor reactivity and decreasing vasodilator reactivity of cerebral arteries, decreasing vasoconstrictor and vasodilator reactivity of carotid and abdominal aortic arteries, decreasing vasoconstrictor reactivity and increasing vasodilator reactivity of pulmonary arteries, decreasing vasoconstrictor reactivity of mesenteric arteries and veins and lower extremity arteries. In addition, microgravity can promote the growth of vascular endothelial cells in the large vessels and inhibit the growth of microvascular endothelial cells. This paper summarized the research progress in the effects of microgravity on blood vessels and vascular endothelial cells.

To explore novel antifatigue agents targeting with AMPA receptor, 10 compounds were synthesized and their structures were confirmed by 1H NMR, ESI-MS and elemental analysis. 1-BCP was treated as the leading compound. The antifatigue activities were evaluated by weight-loaded forced swimming test, and the AMPA receptor binding affinities were tested with radioligand receptor binding assays. The results unveiled that 5b appeared to possess potent antifatigue activities and high affinity with AMPA receptor, which deserved further studies.

Osteoporosis treatment always aimed at keeping the balance of bone formation and bone resorption. Recently, prenyl group in natural products has been proposed as an active group to enhance the osteogenesis process. Osthole has both the prenyl group and bone-protective activities, but the relationship is still unknown. In this study we found that osthole exerted a potent ability to promote proliferation and osteogenic function of rat bone marrow stromal cells and osteoblasts, including improved cell viability, alkaline phosphatase activity, enhanced secretion of collagen-I, bone morphogenetic protein-2, osteocalcin and osteopontin, stimulated mRNA expression of insulin-like growth factor-1, runt-related transcription factor-2, osterix, OPG (osteoprotegerin), RANKL (receptor activator for nuclear factor-κB ligand), and the ratio of OPG/RANKL, as well as increasing the formation of mineralized nodules. However, 7-methoxycoumarin had no obvious effects. Osthole also inhibited osteoclastic bone resorption to a greater extent than 7-methoxycoumarin, as shown by a lower tartrate-resistant acid phosphatase activity and lower number and smaller area of resorption pits. Our findings demonstrate that osthole could be a potential agent to stimulate bone formation and inhibit bone resorption, and the prenyl group plays an important role in these bone-protective effects.

Radix Scutellariae (RS), a medicinal herb, is extensively employed in traditional Chinese medicines and modern herbal prescriptions. Two major flavonoids in RS were known to induce osteoblastic differentiation and inhibit osteoclast differentiation, respectively. This study aimed to investigate the effect of Radix Scutellariae extract (RSE) against bone loss induced by mechanical inactivity or weightlessness. A hindlimb unloading tail-suspended rat model (TS) was established to determine the effect of RSE on bone mineral density and bone microarchitecture. Treatment of RSE at 50 mg/kg/day and alendronate (ALE) at 2 mg/kg/day as positive control for 42 days significantly increased the bone mineral density and mechanical strength compared with TS group. Enhanced bone turnover markers by TS treatment were attenuated by RSE and ALE administration. Deterioration of bone trabecula induced by TS was prevented. Moreover, both treatments counteracted the reduction of bone volume fraction, trabecular thickness and number, and connectivity density. In conclusion, RSE was demonstrated for the first time to prevent osteoporosis induced by TS treatment, which suggests the potential application of RSE in the treatment of disuse-induced osteoporosis.

Many chemotherapeutic agents induce apoptosis via a p53-dependent pathway. However, up to 50% of human cancers have p53 mutation and loss of p53 function. Histone deacetylase inhibitors (HDACIs) are emerging as a potentially important new class of anticancer agents. Here, we report that, Trichostatin A (TSA), a pan-HDAC inhibitor, could induce G2/M cell cycle arrest and apoptosis in both colorectal cancer cell lines with wild-type p53 (HT116 cells) and mutant p53 (HT29 cells), although HCT116 cells had more apoptotic cells than HT29 cells. TSA induces apoptosis in both cell lines via the mitochondrial pathway as indicated by decrease of the mitochondrial membrane potential (MMP) and activation of caspase-3. Additionally, TSA induces expression of the pro-apoptotic protein Bax and decreases the expression of the anti-apoptotic proteins Bcl-2 and Bcl-xL in both cell lines. Bax knockdown by siRNA significantly impaired TSA-induced apoptosis in both cell lines. These data suggest that TSA induces G2/M cell cycle arrest and Bax-dependent apoptosis in colorectal cancer cells (HCT116 cells and HT29 cells) by both p53-dependent and -independent mechanisms. However, cells with normal p53 function are more sensitive to TSA-induced apoptosis.

Radix Astragalus has been identified to exert beneficial effects in preventing postmenopausal bone loss. However, the active ingredients and mechanism of action remain unknown. In this study, we examined the effect of Astragaloside II (AST II), which is a monomer of Astragalus saponin, on the viability, proliferation, differentiation and maturation of rat primary osteoblasts, as well as its relevant molecular mechanism. We found that AST II exhibits a significant induction of proliferation, differentiation and mineralization in primary osteoblasts. AST II stimulates osteoblast differentiation at various stages, from early to late stage of differentiated osteoblasts. Furthermore, induction of differentiation by AST II is associated with increased expression of bone morphogenetic protein-2 (BMP-2), activation of Smad1/5/8, ERK1/2 and p38, and increased expression of core-binding factor 1 (Cbfa1)/Runx2. BMP antagonist (Noggin) blocks the effect of AST II on cell differentiation, and Smad1/5/8, p38, Cbfa1 expression, but only partly decreases ERK1/2 activation. This indicates that BMP-2 is essential in AST II-mediated osteoblast differentiation and Smad1/5/8, p38, Cbfa1 activation, and is partly involved in ERK1/2 activation. In conclusion, although in vivo studies are required in the future, as a phyto-saponin of Radix Astragalus, AST II may become a novel candidate that is beneficial for stimulating the osteoblastic activity resulting in bone formation, which has not been recognized and reported previously.

Aim: To investigate the role of membrane cholesterol in TNFR1-mediated signal transduction in osteoblastic MC3T3 cells., Methods: MCD binds cholesterol specifically and was commonly used to deplete cholesterol from cell plasma membrane. MC3T3 cells were serum-starved for 22 h, treated with MCD (10 g/L) for 60 min followed by TNF-α (10 μg/L) for 0, 5, 10, 15 or 30 min, or TNF-α plus CHX (10 mg/L) for 4 h to induce apoptosis, then TNFR1-mediated IκBα degradation, phosphorylation of AKT, ERK or p38, and processing of caspase-3 were analyzed by using SDS-PAGE/Western blotting method., Results: MC3T3 cell membrane cholesterol level was reduced to 35% within 60 min by MCD (10 g/L). Reduction of MC3T3 cell surface cholesterol dramatically inhibited TNFR1-mediated AKT phosphorylation, while did not affect the degradation of IκBα, activation of ERK or p38, and processing of caspase-3 induced by TNF-α., Conclusion: Cholesterol depletion can destruct lipid rafts; therefore our results suggest that lipid raft is essential for TNFR1-mediated AKT phosphorylation, but is dispensable for TNFR1-mediated degradation of IκBα, activation of ERK or p38 and processing of caspase-3.

Docetaxel is a member of the taxane anti-microtubule class of chemotherapeutic agents, which are currently widely used in clinical cancer therapy. However, the anti-tumor mechanisms of docetaxel are not fully understood. Herein we show that docetaxel induces dose-dependent apoptosis in non-small cell lung cancer A549 cells, as detected by Annexin-V positive cells and PARP cleavage, which is via mitochondrial pathway and dependent on caspase-3 activation. Our study on the mechanisms confirms that docetaxel induces dose-dependent accumulation of cells in M phase and acetylation of α-tubulin, marker of tubulin stablization. Furthermore, docetaxel induces replication-dependent γ-H2AX formation which plays a crucial role in docetaxel-triggered apoptosis. The DNA polymerase inhibitor aphidicolin dose-dependently prevents docetaxel-induced γ-H2AX formation, as well as apoptosis. Notably, 0.6 µM APC almost completely blocked docetaxel-induced γ-H2AX formation and apoptosis. In addition, wortmannin pretreatment caused elevated γ-H2AX level, which was accompanied with increased apoptosis. This effect was due to the inhibition of DNA repair process by wortmannin, as down regulation of p21Waf1/Cip1 and DNA repair proteins such as Ku70, Ku80, DNA-PKcs and Rad50, were detected. These data show, for the first time, that the induction of apoptosis by docetaxel requires DNA replication, and replication-mediated DSBs are critical triggers of docetaxel-induced apoptosis.

Calcium channel blockers (CCBs) are widely used in the therapy of cardiovascular diseases. Recent studies have shown that several CCBs exerted distinct anti-inflammatory effect in myocardial dysfunction models. The purpose of the present study was to evaluate therapeutic effect and possible mechanism of action of amlodipine, one of the widely used CCBs, on rat cardiac dysfunction during sepsis induced by lipopolysaccharide (LPS). Pretreatment of the rats with amlodipine (10 or 30 mg/kg, i.v.) delayed the fall of mean arterial blood pressure caused by LPS. Amlodipine also significantly inhibited the elevation of plasma tumor necrosis factor alpha (TNF-alpha) and decreased levels of inducible nitric oxide synthase (iNOS) in response to LPS challenge. To investigate the mechanism of the action of amlodipine, neonatal rat cardiomyocytes were used as a model. Amlodipine concentration-dependently decreased the release of TNF-alpha and iNOS protein expression, and suppressed the degradation and phosphorylation of inhibitor of kappaB-alpha (IkappaB-alpha) in LPS-activated neonatal rat cardiomyocytes. Further studies revealed that amlodipine markedly activated phosphatidylinositiol 3-kinase (PI3K) and Akt, downstream of the PI3K signal cascade. Application of PI3K inhibitors, wortmannin and LY294002 attenuated the depression of TNF-alpha and iNOS expression by amlodipine in LPS-induced cardiomyocytes. These findings may explain some cardioprotective effects of amlodipine in LPS-mediated sepsis and suggest that the inhibition of TNF-alpha and iNOS expression by amlodipine is, at least in part, dependent on PI3K/Akt signaling pathway.

Histone deacetylase (HDAC) inhibitors (HDIs) play an important role in the regulation of gene expression associated with cell cycle and apoptosis and have emerged as promising anticancer agents. In addition to their intrinsic anticancer properties, some studies have demonstrated that HDIs can modulate cellular responses to ionizing radiation (IR). Here we show evidence that co-treatment with the HDI trichostatin A (TSA) radiosensitizes human non-small cell lung cancer (NSCLC) A549 cells and H1650 cells. Cells were exposed to gamma-irradiation with or without TSA co-treatment. Clonogenic survival was significantly reduced in cells with TSA co-treatment. In A549 cells, TSA enhanced IR-induced accumulation of cells in G(2)/M phase, with upregulated expression of p21(waf1/cip1). In addition, TSA co-treatment caused pronounced apoptosis in irradiated cells, which was accompanied with p21(waf1/cip1) cleavage to a 15 kDa protein. The enhanced apoptotic effect was via mitochondrial pathway, as indicated by the increased dissipation of mitochondrial transmembrane potential (MMP) and release of cytochrome c from the mitochondria to the cytoplasm. Caspase-3 activation was also significantly increased, with accordingly more cleavage of PARP, associated with the repression of X-linked inhibitor of apoptosis protein (XIAP). Furthermore, TSA co-treatment impaired DNA repair capacity after IR by downregulation of Ku70, Ku80 and DNA-PKcs, reflected by enhanced and prolonged expression of gammaH2AX. Taken together, our results demonstrate that TSA acts as a powerful radiosensitizer in NSCLC cells by enhancing G(2)/M cell cycle arrest, promoting apoptosis through multiple pathways and interfering with DNA damage repair processes.

Carbohydrates are proved to be involved in recognition processes, including adhesion between cells, adhesion of cells to the extracellular matrix, and specific recognition of cells by one another. Galectin-3 is a nonenzymatic carbohydrate-binding protein present in mammals, whose conserved carbohydrate-recognition domain preferentially binds to specific carbohydrate structures and plays an important role in biological and biochemical reactions. Substances developed with the specific structure of beta-galactose moieties or their analogues, including chemically modified carbohydrates, functional peptides and modified natural polysaccharide, have been evaluated as potent therapeutic ligands for galectin-3 and showed at different level their ability to interfere with carbohydrate-protein interactions and therefore, inhibit the cell-cell recognition and adhesion processes, which play an important role in tumor growth, progression and metastasis.

The polysaccharide APS-2a was isolated from Angelica sinensis (Oliv.) Diels through water extraction, deprotein, ethanol precipitation and DEAE-sephades A-25 column chromatography respectively,and was further purified by Sephacryl S-400 and Sephadex G-100 column chromatography. The phenol-sulfuric acid assay and Bradford method were used to determine the contents of carbohydrate and protein, respectively. The molecular weight was carried out with high-performance size exclusion chromatography (HPSEC) combined with a differential refractometer detector. The monosaccharide compositions were determined by gas chromatography after complete hydrolysis with acid. The models of mice transplanted sarcoma S-180 were used to study the anti-tumor effects in vivo. Thymus indexes, spleen indexes were determined. The HPSEC result showed the APS-2a was a single homogeneous component and its weight average molecular weight was 7.4 x 10(5) Da. The monosaccharide composition of APS-2a was glucose, galactose, arabinose, rhamnose, galcturonic acid. Furthermore, APS-2a (20.50 mg/kg) could inhibit the proliferation of tumor cells in mice transplanted S-180. The thymus indexes and spleen indexes in the groups treated with APS-2a were higher than control group.

Objective: To investigate the anti-oxidative effect of Angelica polysaccharide sulphate( APS)., Methods: The Hela cells were cultured conventionally. Then APS was added and cultured together in different concentration for 24h followed by a oxidative injury with H2O2 or UV irradiation. The anti-oxidative effects of APS were detected as follow: cell viability was measured by MTT assay; colorimetric analysis was used to determine SOD activity, GSH and MDA level in cytoplasm., Results: Treatment of H2O2 or UV irradiation significantly decreased cell viability, GSH and SOD activity in cytoplasm,while increased MDA in cytoplasm. At the range of 0. 3 -100microg/ml, APS significantly increased cell viabilty, GSH and SOD activity,while decreased MDA in a dose dependent manner( P < 0. 05 or P <0.01)., Conclusion: APS has anti-oxidative effect,which may be one of its anti-AIDS mechanisms.

Aim: To study the biliary excretion of genistein and its metabolite at different doses in rats., Methods: Suspended in 0.5% CMC-Na solution, genistein was orally administered to rats at the dose of 6.25, 12.5 and 50 mg x kg(-1), separately. At various time intervals, the bile was collected. The bile was treated with beta-glucuronidase. The genistein in bile was extracted twice by vortexing with 2.0 mL mixture of methyl tert-tubtyl ether and pentane (8:2). The organic phase was removed into the tubes and then evaporated in ventilation cabinet. The residue was dissolved in 50 microL of methanol. Twenty microL solution was drawn and detected by high-performance liquid chromatography., Results: The accumulative biliary excretion of genistein was (42.56 +/- 6.54) , (75.17 +/- 18.87) and (126.60 +/- 34.78) microg at the dose of 6.25, 12.5 and 50 mg x kg(-1), respectively. The total drug (genistein plus glucuronidated genistein) excreted from bile was (108.46 +/- 35.23), (423.46 +/- 158.31) and ( 853.74 +/- 320. 84) microg, and the ratio of glucuronidated genistein was 60.76% , 82.25% and 85.17% at the dose of 6.25, 12.5 and 50 mg x kg(-1), respectively., Conclusion: The genistein was excreted mainly in the form of glucuronidated genistein in rat bile. The genistein and glucuronidated genistein were excreted in a nonlinear dose-dependent manner.

Aim: To investigate the effects of Angelica sinensis polysaccharide fraction AP-3 on IL-2 and IFN-gamma induction and its further immunomodulatory feature., Methods: The percentage of CD4+ lymphocyte was detected by flow cytometric method, the production of IL-2 and IFN-gamma in cell culture supernatant were determined by ELISA, mRNA expressions of IL-2 and IFN-gamma cytokines were detected by RT-PCR., Results: At the range of 0. 6 - 2 micromol x L(-1), AP-3 significantly enhanced the percentage of CD4+ lymphocytes in total splenocytes. At the range of 2 - 6 micromol x L(-1), the treatment of AP-3 augmented both productions of IL-2 in cell culture supernatant and cell IL-2 mRNA transcription level in a time and dose dependent manner. While in the case of IFN-gamma, AP-3 stimulated at early time after exposure but down-regulated thereafter., Conclusion: Angelica sinensis polysaccharide could regulate the immune response through upregulating IL-2, IFN-gamma expression and activating Th1 cell.

Aim: To investigate the effects of Angelica sinensis polysaccharide (AP) on cell-mediated immunity., Methods: The lymphocyte proliferation was tested by MTT colormetry; T lymphocyte fraction were separated with immune magnetic beads and the change of CD4(+) T lymphocyte percentage was detected by flow cytometry., Results: In the range of 30-300 mg/L, the total AP and its three fractions, ie AP-0, AP-1, AP-2, AP-3, directly stimulated the proliferation of murine splenocytes, and AP-3 significantly enhanced the proliferation of lymphocytes in MLR and T lymphocytes. The percentage of CD4(+) T lymphocytes in total splenocytes was also enhanced by AP-3., Conclusion: AP can promote the cell-mediated immunity with T lymphocytes as one of its target cells.

Aim: To study the metabolic kinetics of MN9202 in Beagle dog liver microsome., Methods: Beagle dog liver microsomes were prepared by using ultracentrifuge method. After incubating 0.4 micromol x L(-1) MN9202 with 1 g x L(-1) microsomes for 30 min at 37 degrees C, the reaction was terminated by adding 0.5 mL alkalization. The RP-HPLC was used to determine the drug in the incubation mixture. The Michaelis-Menten parameters Km, and Vmax in Beagle dog liver microsomes were initially estimated by analyzing Lineweave-Brurk plot. Various selective CYP inhibitors were used to investigate their inhibitory effect on the metabolism of MN9202., Results: The Km, Vmax and CLint of MN9202 were (22.6 +/- 8.0) micromol x L(-1), (0.54 +/- 0.17) micromol x g(-1) x min(-1) and (0.0242 +/- 0.0009) L x g(-1) x min(-1), respectively. The metabolism of MN9202 was significantly inhibited by ketoconazole (Ket) and troleandomycin (Tro) in Beagle dog liver microsomes. Tranylcypromine (Tra) could inhibit the metabolism of drug as well. While other inhibitors showed little inhibitory effect on the metabolism of MN9202., Conclusion: It was shown that CYP3A and CYP2C19 were involved in MN9202 metabolism. The inhibitors of human CYP3A and CYP2C19 may have potential interaction with MN9202, and this can reduce the metabolism rate and increase the toxicity of MN9202.

Aim: To study the pharmacokinetics of genistein at different doses in Beagle dogs., Methods: Suspended in 0.5% CMC-Na solution, genistein was orally administered to Beagle dogs at doses of 2.67, 5.34 and 10.68 mg.kg(-1). At various time intervals, 1.5 mL of blood was drawn from the femoral vein of dogs in their front legs. The plasma was treated with beta-glucuronidase. The genistein in plasma was extracted twice by vortexing with 2.0 mL mixture of methyl tert-tubtyl ether and pentane (v/v = 8:2). The organic phase was removed into the tubes and then evaporated in ventilation cabinet. The residue was dissolved in 50 microL of methanol. 20 microL solution was drawn and detected by high-performance liquid chromatography. The pharmacokinetic parameters were calculated by 3P97 software., Results: The plasma drug concentration-time data were fitted to the two-compartment model. When the dose was 2.67 mg.kg(-1), the MRT and AUC of parent compound were 52.9 min and 6.7 mg.min. L(-1), respectively. When the dose rose to 5.34 mg.kg(-1), the MRT and AUC of parent compound became 224.8 min and 26.1 mg.min.L(-1), respectively. However, when the dose increased to 10.68 mg .kg(-1), the MRT and AUC of parent compound increased to 267.7 min and 33.2 mg.min L(-1), respectively. The AUC of glucuronidated genistein was 33.9, 70.1 and 140.5 mg.min.L(-1) at the dose of 2.67, 5.34 and 10.68 mg.kg(-1), respectively., Conclusion: Due to significant first pass metabolism, the drug was mainly existed in the form of glucuronidated genistein in the plasma. With the increase of dose, the absorption of genistein became saturated and the half life prolonged.

Aim: To describe the effect of Rheum tanguticum polysaccharide (RTP) on hydrogen peroxide-induced human intestinal epithelial cell injury., Methods: Hydrogen peroxide (100 micromol/L) was introduced to induce human intestinal epithelial cell injury. Cells were pretreated with RTP (30,100,300 microg/mL) for 24 h before exposure to hydrogen peroxide. Cell viability was detected by MTT assay and morphological observation. Acridine orange staining and flow cytometry were performed to assess cell apoptosis. Lactate dehydrogenase (LDH) activity, production of malondialdehyde (MDA) and superoxide dismutase (SOD) activity were measured by spectrophotometry with corresponding assay kits., Results: Following exposure to H(2)O(2), a marked decrease in cell survival and SOD activity, increased production of MDA, LDH leakage and cell apoptosis were found. Pretreatment of the cells with RTP could significantly elevate cell survival, SOD activity and decrease the level of MDA, LDH activity and cell apoptosis., Conclusion: RTP may have cytoprotective and anti-oxidant effects against H(2)O(2)-induced intestinal epithelial cell injury by inhibiting cell apoptosis and necrosis. This might be one of the possible mechanisms of RTP for the treatment of ulcerative colitis in rats.

Aim: To study whether PC-407 [4-[5-naphthyl-3- (trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide] inhibits cell viability and induces apoptosis in human colon cancer SW-1116 cells., Methods: Inhibition of SW-1116 proliferation was measured by MTT assay. Morphological assessment of apoptosis was performed with fluorescence microscope and electron microscope. DNA fragmentation was visualized by agarose gel electrophoresis. The amount of apoptotic cells was measured by flow cytometry., Results: PC-407 inhibited SW-1116 cell proliferation in a concentration-dependent manner after 3 d of treatment, and the IC(50) for PC-407 inhibition of cell number was 16.67+/-0.17 micromol/L. After incubation of SW-1116 cells with PC-407 20 micromol/L for 24 h, morphological changes of typical apoptosis were observed by AO/EB staining or transmission electron microscopy. Flow cytometry analysis showed that PC-407 induced apoptosis in SW-1116 cells in a time- and concentration-dependent manner. The agarose gel electrophoresis of DNA revealed a ladder pattern 48 h later., Conclusion: PC-407 inhibited proliferation and induced apoptosis in the human colon cancer SW-1116 cell line.

Aim: To observe the effect of Angelica polysaccharides on effector molecule production by peritoneal macrophages., Methods: Macrophages were isolated from the peritoneal cavity of BALB/c mice and the primary culture was performed. MTT colorimetry and spectrophotometry were used to examine the effects of Angelica polysaccharides on the releases of effector molecules, such as nitric oxide(NO), tumor necrosis factor-alpha(TNF-alpha), and reactive oxygen species(ROS) as well as inducible nitric oxide synthase (iNOS) and lysozyme(LSZ) activity by peritoneal macrophages., Results: Angelica polysaccharides could promote the releases of NO, TNF-alpha and ROS from macrophages and improved iNOS and LSZ activities in macrophages. However, Angelica polysaccharides had no direct cytotoxicity to tumor cells, but the cultural supernatant of macrophages cocultured with Angelica polysaccharides could kill L929 cells., Conclusion: Angelica polysaccharides can promote the releases of NO, TNF-alpha and ROS by macrophages. Angelica polysaccharides may indirectly play the role of anti-tumors through increased TNF-alpha production by macrophages.

To investigate the protective effects of nitric oxide (NO) on cardiomyocytes against hydrogen peroxide (H2O2)-induced injury, cultured neonatal cardiomyocytes were divided into three groups: (1) normal group; (2) H2O2 group: cells were treated with H2O2 (0.1 mmol/L) for 4 h; (3) SNAP+ H2O2 group: cells were pretreated with NO donor S-nitroso-N-acetyl-1,1-penicillamine (SNAP, 0.5 mmol/L) 10 min before H2O2 treatment. Colorimetric assay was used to detect cell viability and lactate dehydrogenase (LDH) activity to evaluate cell injury. Apoptotic rate of cardiomyocytes were determined by flow cytometer. Superoxide dismutase (SOD) activity and malonaldehyde (MDA) content were measured by colorimetric assay to evaluate cell antioxidant ability. Intracellular calcium was tested by laser confocal microscopy. The results showed that after treatment with H2O2, cell viability was significantly reduced to 58.3+/-7.6% compared with normal group (93.1+/-6.2 %). LDH activity and apoptotic rates were 1580.5+/-186.7 U/L and 26.4+/-5.7% respectively, significantly higher than that of normal group (631.4+/-75.6 U/L and 0). SNAP pretreatment markedly improved cell viability to 79.7+/-9.3% and reduced LDH activity and apoptotic rates to 957.8+/-110.9 U/L and 9.1+/-3.3%, respectively. Cells treated with H2O2 had a lower SOD activity of 14.73+/-1.68 NU/ml and a higher MDA content of (15.35+/-3.49) micromol/L compared with normal cells (19.67+/-0.85 NU/ml) and (6.95+/-0.83 micromol/L), respectively. Cells with SNAP pretreatment had a higher SOD activity of 21.36+/-3.11 NU/ml and a lower MDA content of 9.12+/-1.47 micromol/L compared with H2O2 group. Intracellular calcium content was reduced by SNAP administration while enhanced by H2O2. Pretreatment with SNAP could antagonize the effect of H2O2 of accelerating intracellular calcium content. Based on the results observed, it is concluded that NO donor SNAP may protect cardiomyocytes from being injured by H2O2. The underlying mechanisms may include improving cell antioxidant ability and reducing intracellular calcium overload.

Aim: To study the pharmacokinetics of m-nifedipine (m-Nif) in Beagle dogs., Methods: The Beagle dogs were divided into two groups. m-Nif was intravenously administered to the Beagle dogs in group 1 at the dose of 0. 288 mg x kg(-1), and it was orally administered to the Beagle dogs in group 2, 3 and 4 at the dose of 1.152, 3.456 and 10.370 mg x kg(-1), respectively. m-Nif in plasma was detected by reversed phase high performance liquid chromatography. The pharmacokinetic parameters were calculated by 3P97 software., Results: When m-Nif was intravenously administered, the plasma concentration-time curve was fit to a two-compartment model and T1/2beta was 117 min. When m-Nif was orally administered, the plasma concentration-time curve was fit to a one-compartment model. T1/2 (Ke) and Cmax were 147 min and 20 microg x L(-1); at the low dose of 1.152 mg x kg(-1). T1/2 (Ke) was 122 min and Cmax was 36 microg x L(-1) at the middle dose of 3.456 mg x kg(-1). T1/2 (Ke) was 144 min and Cmax was 69 microg x L(-1) at the high dose of 10.37 mg x kg(-1), respectively., Conclusion: It was showed that the speed of elimination of m-Nif was high in Beagle dogs. The absolute bioavailability of m-Nif given orally was very low.

Objective: To investigate the effect of puerarin on the neural function and the histopathological changes after ischemic spinal cord injury in rabbits., Methods: Thirty male New Zealand white rabbits were randomly divided into three groups as follows: puerarin group (n=10) receiving intravenous infusion of 30 mg/kg puerarin for 10 minutes, control group (n=10) receiving intravenous infusion of the same volume of normal saline as puerarin for 10 minutes, and sham operation group (n=10) undergoing only the surgical exposure of the abdominal aorta. Temporary spinal cord ischemia was induced by infrarenal aortic occlusion for 20 minutes and followed by reperfusion. The neural status was scored with the Tarlov criteria at 8, 12, 24 and 48 hours after reperfusion. All the animals were killed at 48 hours after reperfusion and the spinal cords (L5) were removed immediately for histopathological study., Results: The neural function scores at 8, 12, 24 and 48 hours after reperfusion were higher in the puerarin group and sham operation group than those in the control group (P<0.05). More normal motor neurons in the anterior horn of spinal cord were present in the puerarin group and sham operation group than those in the control group (P<0.01). There was a strong correlation between the final neural function scores and the number of normal motor neurons in the anterior horn of spinal cord (r=0.839, P<0.01)., Conclusions: Puerarin can significantly ameliorate the neural function and the histopathological damages after transient spinal cord ischemia in rabbits.

Aim: To examine if estrogen can affect the immune response at the dendritic cells (DCs) level in rats with experimental autoimmune encephalomyelitis (EAE)., Methods: Lewis rats were immunized with inoculum containing MBP(68-86). DCs were derived from spleen monocytes of EAE rats with IL-4 and GM-CSF in presence of 17 beta-estradiol (E2). Nitric oxide (NO) was detected by Griess reagent. The surface markers and cytokines production of DCs were shown by flow cytometry. DCs were cocultured with MBP-specific T cells, [(3)H]-TdR incoportation was used to reveal the antigen presentability, the supernatant of the coculture were collected to examine the cytokines secretion by ELISA., Results: E2 activated DCs by accelerating the maturation process characterized by upregulation of MHC II and costimulating molecule B7-1, B7-2, drastic high expression of CD40. IFN-kappa-producing DCs were also elevated without any alteration of IL-10. Estradiol-treated DCs (E2-DCs) secreted more NO in the culture supernatant. By contrast, E2-DCs showed decreased antigen presentation ability with reduced secretion of IFN-kappa but no alteration of IL-10 in the coculture with T cells., Conclusion: Estrogen can affect the differentiation, maturation and function of DCs from EAE rats, which may be attributed to its protection against EAE and the remission of multiple sclerosis patients in pregnancy.

Aim: Stress induces gastric ulceration in human and experimental animals. People tend to smoke more cigarettes when under stress. Nitric oxide (NO) and nicotine have opposing effects on gastric integrity. The present study examined the possible therapeutic benefit of NO in nicotine-treated rats with stress-induced gastric ulceration., Methods: Rats drank a nicotine solution while control rats drank tap water for 20 days. The alkoloid was then replaced by water with or without supplementation of isosorbide dinitrate (NO donor) for an additional 10 days. Isosorbide dinitrate was given twice shortly before experiments (acute) or three times daily by oral gavages for 10 days after the rats stopped drinking nicotine solution. At the end of experiments, ulcer index, gastric adhesion mucus content and MPO activity were measured and analysed., Results: Nicotine treatment decreased gastric mucus content and intensified stress-induced gastric ulcer. A higher ulcer index persisted even after the rats stopped drinking nicotine solution for 10 days. Acute NO donor showed no benefit on both mucus and ulcer index in nicotine treatment or/and stress condition. Chronic NO donor treatment reversed the worsening action of nicotine in stomach. Stress increased gastric mucosal myeloperoxidase (MPO) activity, which was antagonized by chronic NO treatment. However, nicotine was unlikely to change mucosal MPO activity., Conclusion: The intensifying action of nicotine on stress-induced gastric ulceration persists for 10 days after cessation. Nicotine treatment significantly decreases gastric mucus content that can be restored by chronic NO donor treatment. The present study suggests that NO antagonizes the ulcerogenic action of nicotine through a cytoprotective way.

Aim: To study the effects of tumor necrosis factor-alpha (TNF-alpha) on calcium movement in rat ventricular myocytes., Methods: Intracellular free Ca2+ concentration was measured with calcium fluorescent probe Fluo-3/AM and laser confocal microscope. L-type calcium current (ICa,L) was recorded with the whole-cell configuration of the patch-clamp techniques., Results: At 2, 20 and 200 microg/L, TNF-alpha was found to increase intracellular free Ca2+ concentration in a dose-dependent manner illustrated by the increment of calcium fluorescence density with laser confocal microscope. Nicardipine 0.5 micromol/L slightly attenuated TNF-alpha-induced response. When the cardiac myocytes were exposed to caffeine (100 mmol/L) for 30 min, TNF-alpha failed to induce any change of intracellular free calcium. However, it was found that TNF-alpha inhibited I(Ca,L) in whole-cell patch-clamp experiments. At 2, 20, and 200 microg/L, TNF-alpha decreased peak I(Ca,L) by 3.9 % (-5.1 pA/pF+/-0.3 pA/pF vs -4.9 pA/pF+/-0.2 pA/pF, n=9, P>0.05), 15.7 % (-5.1 pA/pF+/-0.3 pA/pF vs -4.3 pA/pF+/-0.3 pA/pF, n=9, P<0.05) and 19.6 % (-5.1 pA/pF+/-0.3 pA/pF vs -4.1 pA/pF+/-0.4 pA/pF, n=9, P<0.01), respectively. It shifted the steady-state inactivation curve of I(Ca,L) to the left (V1/2 shifted from -28.7 mV+/-0.3 mV to -37.8 mV+/-1.4 mV, n=7, P<0.05), while it took no effects on steady-state activation and recovery from inactivation., Conclusion: TNF-alpha inhibited I(Ca,L) in rat ventricular myocytes, while increasing the intercellular free Ca2+ level due to the release of Ca2+ from intracellular stores.

Objective: To evaluate protective effects of Rheum tanguticum polysaccharides (RTP) on traumatic brain injury (TBI) in rats., Method: The polysaccharides (RTP) were extracted from Tanguficum Maxim. 120 rats were divided into 15 groups, with 8 rats in each group. RTP at 100, 200 and 400 mg x kg(-1) were administrated orally once a day for five days, and model of brain injury was made by dropping weight method., Result: RTP reduced water content and malondialdehyde (MDA) levels, and increased total SOD activity and Na+-K+ ATPase activity after injuried., Conclusion: The polysaccharides may be one of the effective comptents in Rheum tanguticum, showing significant neuroprotective effects.

Aim: To study the effects of Rheum tanguticum polysaccharide(-1) (RTP(-1)) on ulcerative colitis in rats induced by 2, 4, 6-trinitrophene sulphonic acid (TNBS) and their possible mechanism., Methods: RTP1 (200 mg.kg(-1), i.g.) extracted from Rheum tanguticum Maxim. ex Regel was administrated to rats with colitis induced by TNBS for 5 d, 7 d, 10 d and 14 d, respectively. The effects of RTP1 and dexamethasone (DX, 0.2 mg.kg(-1), i.g.) were contrastively investigated. The MPO level and SOD activity were determined by chromatometry. The expansion and protein expression of CD4+T lymphocytes isolated from colon mucosae and mesenteric lymph nodes of colitis rats were performed by immunohistochemical analysis and Western-blot methods., Results: Treatments of RTP1 (200 mg.kg(-1), i.g.) significantly reduced diarrhea, mortality, colon mass, ulcer areas and MPO level in colon mucosae on days 5, 7, 10 and 14 (5.2+/-1.4, 5.4+/-0.7, 5.2+/-1.8, P<0.05. 3.4+/-0.8, P<0.01. 16.1+/-12.1, P<0.01. 31.8+/-8.6, 17.7+/-5.3, 12.7+/-4.1, P<0.05). The effects of RTP1 were similar to those noted above in DX group, but there were no immunosuppressive effects of DX in RTP(-1) group, such as body mass loss, thymus and spleen atrophy. The decreased number and down-regulated protein levels of CD4+T cells isolated from the colon of colitis rats treated with RTP1 were found., Conclusion: RTP1 shows significantly protective effects but lower side effects on rats with colitis induced by TNBS. The mechanism may be due to the resistance to over expansion of CD4.

Aim: To study the pharmacokinetics of genistein in Beagle dogs., Methods: Genistein, suspended in 0.5% CMC-Na solution, was orally administered to Beagle dogs at the dose of 5.34 mg.kg-1. At various time intervals, 1.5 mL of blood was drawn from the vein of dogs in their front legs. At the same time, urine and feces were collected. After the collection, the feces were homogenized with physiological saline (to 1 g feces, 10 mL physiological saline were added). The genistein in plasma, urine and homogenized feces was extracted twice by vortexing with 2.0 mL mixture of methyl tert-butyl ether and pentane (8:2). The organic phase was transferred into tubes and evaporated in ventilation cabinet. The residue was dissolved in 50 microL of methanol and 20 microL of the solution was drawn and detected by high-performance liquid chromatography. The pharmacokinetic parameter was calculated by 3P97 software., Results: The plasma concentration-time curve was fitted to a one-open-compartment model. The peak time was 0.29 h, and the elimination half-life was 0.52 h. After genistein was administered, 10.79% of genistein were excreted from urine and 21.55% from feces within 24 h. It was also found that 13.00% genistein were excreted from urine and 52.46% from feces within 60 h., Conclusion: It showed that the speed of absorption and elimination of genistein was high in Beagle dog, and genistein was mainly excreted in the form of parent compound in urine and feces.

Aim: To investigate the in vivo anti-tumor effects of total polysaccharide (AP-0) isolated from Angelica sinensis (Oliv.) Diels (Danggui) on mice and the in vitro inhibitory effects of AP-0 and the sub-constituents (AP-1, AP-2 and AP-3) separated from AP-0 on invasion and metastasis of human hepatocellular carcinoma., Methods: Three kinds of murine tumor models in vivo, sarcoma 180 (S180), leukemia L1210 and Ehrlich ascitic cancer (EAC) were employed to investigate the anti-tumor effects of AP-0. For each kind of tumor model, three experimental groups were respectively given AP-0 at doses of 30, 100 and 300 mg/kg by ip once a day for 10 days. Positive control groups were respectively given Cy at a dose of 30 mg/kg for S180 and leukemia L1210, and 5-FU at a dose of 20 mg/kg for EAC. On d 11, mice bearing S180 were sacrificed and the masses of tumors, spleens and thymus weighed. The average living days of mice bearing EAC and of mice bearing L1210 were observed, and the rates of life prolongation of each treatment were calculated, respectively. The inhibitory effects of APs on hepatoma invasion and metastasis in vitro were investigated by employing human hepatocellular carcinoma cell line (HHCC) with the Matrigel invasion chamber, adhesion to extracellular matrix and chemotatic migration tests, respectively., Results: AP-0 had no obviously inhibitory effect on the growth of S180, but it could significantly decrease the thymus weights of the mice bearing S180. AP-0 could significantly reduce the production of ascitic liquids and prolong the life of mice bearing EAC. AP-0 could also increase the survival time of mice bearing L1210. AP-0 and AP-2 had significantly inhibitory effects on the invasion of HHCC into the Matrigel reconstituted basement membrane with the inhibitory rates of 56.4 % and 68.3 %, respectively. AP-0, AP-1, AP-2 and AP-3 could influence the adhesion of HHCC to extracellular matrix proteins (Matrigel and fibronectin) at different degrees, among them only AP-3 had significant blocking effect on the adhesion of HHCC to fibronectin with an inhibitory rate of 30.3 %. AP-0, AP-1 and AP-3 could partially inhibit the chemotactic migration abilities of HHCC., Conclusion: The experimental findings suggest that the total polysaccharide of Angelica sinensis (Oliv.) Diels (Chinese Danggui) possesses anti-tumor effects on experimental tumor models in vivo and inhibitory effects on invasion and metastasis of hepatocellular carcinoma cells in vitro.

Aim: To evaluate the effects of molecular weight of dextran on drug-release of conjugate in vitro by screening colon-specific conjugates., Methods: The conjugates, synthesized with different molecular-weight dextran and dexamethasone, were incubated in the contents of different parts of rat gastrointestinal tract at 37 degrees C. The release of dexamethasone(Dex) and dexamethasonehemisuccinate was determined by HPLC. The mobile phase consisted of 35% acetonitrile and 65% trisodium citrate (50 mmol.L-1, adjusted to pH 4.1 with phosphoric acid)., Results: There was no release of dexamethasone or dexamethasonehemisuccinate from conjugates in the stomach contents. The amount of Dex (including dexamethasonehemisuccinate) released from DexD26 in the contents of colon and cecum was shown to be 4.0 times higher than that released in the contents of proximal and distal small intestine while the amount of Dex (including dexamethasonehemisuccinate) released from DexD50 was shown to be 3.6 times higher. The amount of Dex (including dexamethasonehemisuccinate) released from DexD2 in the contents of colon and cecum and from DexD7.6 were 2.0 times and 1.9 times higher, respectively, than that released in contents of proximal and distal small intestine., Conclusion: The molecular weight of dextran showed marked effect on drug-release of the conjugate in vitro, and the conjugates with larger molecular-weight dextran have great potential in colon-specific delivery of dexamethasone.

Objective: To evaluate the effects of Tanguticum Maxim polysaccharide (TMP-1) on TNBS-induced colitis in rats., Method: Rats with TNBS/ethanol-induced colitis were used and treated with TMP-1 and dexamethasone (DX). Seventy-two rats, including animals with TNBS-induced colitis, were treated with saline, TMP-1 (100, 200, 400 mg.kg-1) and DX. White blood cells were counted on the fifth day and the rats were killed by ether on the sixth day. SOD activity in serum, MPO and SOD activity of colonic tissue were measured., Result: The remarkable effects of TMP-1 at dosage of 200, 400 mg.kg-1 on TNBS-induced colitis were observed. The ulcerative area was diminished and weight of colon was reduced. White blood cell population was reduced, SOD activity in serum and SOD activity of colon tissue were remarkably increased, and, MPO activity of colonic tissue was reduced., Conclusion: TMP-1 has significant effects on TNBS-induced colitis in rats with lower side effects, which suggests the effective component of rhubarb on colitis perhaps is TMP. The mechanism of the actions of TMP may relate to its antiflammation, antioxidation and immunoloregulation.

Aim: To study the effects of serotonin (5-hydroxy-tryptamine, 5-HT) on transient outward potassium current (I(to)) and elucidate its mechanism in rat ventricular myocytes., Methods: I(to) was recorded using the conventional whole cell patch-clamp techniques., Results: I(to) density in normal myocytes was similar to that in norepinephrine-induced hypertrophic myocytes. 5-HT depressed I(to) in a concentration-dependent manner with the half-maximal inhibitory concentration of (40+/-5) micromol/L and (38+/-7) micromol/L in normal and hypertrophic ventricular myocytes respectively. Mianserin (5-HT2 receptor antagonist), compound 48/80 (phospholipase C antagonist), and chelerythrine chloride (protein kinase C antagonist) reversed the inhibitory effects of 5-HT on I(to), while phorbol 12-myristate 13-acetate (protein kinase C agonist) enhanced the inhibitory effect of 5-HT on I(to) in normal myocytes., Conclusion: 5-HT markedly inhibits I(to) in rat ventricular myocytes. The putative signal pathway is that 5-HT activates phospholipase C, which causes inositol phospholipid hydrolysis. The activation of downstream signal molecule, protein kinase C, phosphorates substrate target proteins, which leads to inhibition of I(to) in ventricular myocytes.

Aim: To study the effects of 5-hydroxytryptamine (5-HT) on L-type calcium current (ICa) in norepinephrine (NE)-induced hypertrophic ventricular myocytes., Methods: Left ventricular hypertrophy was induced by injecting NE intraperitoneally in rats. The single myocytes were isolated enzymatically from left ventricle. ICa was recorded with the whole-cell configuration of the patch-cl amp technique., Results: (1) The ratio of left heart weight to body weight (LHW/BW) was higher (P < 0.01) in the NE-treated rats compared with the control rats on d 15. LHW/BW was increased 31.8 % in NE-treated rats. (2) ICa was larger in hypertrophic cells than that in normal cells (4.5 p A/pF +/- 0.5 pA/pF vs 3.5 pA/pF +/- 0.3 pA/pF, respectively, at testing potential of 0 mV; P < 0.01). (3) 5-HT (1, 10 micromol/L) increased ICa and decreased the peak current potential from 0 mV to -10 mV in both myocytes. The augmentation of ICa induced by 5-HT was larger in hypertrophic ones. (4) 5-HT did not markedly influence the steady-state activation kinetics. However, 5-HT shifted steady-state inactivation curve with half inactivation voltage V 1/2 changing from -39.5 mV +/- 1.8 mV to -27.8 mV +/- 1.7 m V (P < 0.05), while not changing the voltage responsiveness of calcium channel (slope factor k was not changed markedly)., Conclusion: 5 -HT increased ICa in ventricular myocytes by changing the kinetics of steady-st ate inactivation. A larger alteration of ICa induced by 5-HT i n hypertrophic ventricular myocytes suggests that 5-HT be more prone to induce arrhythmia in hypertrophic heart than in normal one.