Red blood cell distribution width and ischaemic stroke.

The red blood cell distribution width (RDW) is a measure of red blood cell (RBC) size heterogeneity, which is easily calculated by dividing the SD of erythrocyte volumes for the mean corpuscular volume. Recent reporter suggested that, besides haematological diseases and anaemia, many human disorders may be closely associated with the elevated RDW. A literature review has revealed the RDW may be closely related to the development of ischaemic stroke, carotid artery atherosclerosis and cerebral embolism. Higher RDW could independently predict adverse outcomes in patients in these conditions.

Induction of apoptosis in MCF­7 human breast cancer cells by Khz (fusion of Ganoderma lucidum and Polyporus umbellatus mycelium).

Khz (fusion of Ganoderma lucidum and Polyporus umbellatus), isolated from the mycelia of G. lucidum and P. umbellatus, exerts anti­proliferative effects against malignant cells; however, its activity against human breast cancer cells remains to be elucidated. In the present study, cell proliferation was assessed using a 3-(4,5­dimethylthiazol­2­yl)-2,5­diphenyltetrazolium bromide assay, and poptosis was examined using annexin V­propidium iodide staining and flow cytometry. The activation of caspases 7, 8 and 9 were detected in the Khz­treated cells using western blotting. The results demonstrated that Khz increased the intracellular calcium concentration and induced the production of reactive oxygen species in MCF­7 breast cancer cells, as determined using flow cytometry. The results also demonstrated that Khz inhibited cell proliferation and induced apoptosis in the MCF­7 cells. In addition, the mechanism by which Khz induces apoptosis in cancer cells was investigated. Khz induced apoptosis preferentially in transformed cells, with a minimal effect on non­transformed cells, suggesting its potential as an anticancer therapeutic agent. Oxidative stress is associated with apoptotic and non­apoptotic cell death, although pro­oxidative conditions are not a pre­requisite for apoptosis. Assessment of the activation status of caspases 7, 8 and 9 revealed that the levels of cleaved caspases were significantly increased in the cells treated with Khz. It is widely accepted that calcium signaling is important in apoptosis, and the present study observed an increase in [Ca2+]i in response to Khz treatment. The anti­proliferative and pro­apoptotic effects of Khz suggest that this extract may be developed as a potential anticancer agent.

Khz (fusion product of Ganoderma lucidum and Polyporus umbellatus mycelia) induces apoptosis in human colon carcinoma HCT116 cells, accompanied by an increase in reactive oxygen species, activation of caspase 3, and increased intracellular Ca²âº.

Khz (a fusion mycelium of Ganoderma lucidum and Polyporus umbellatus mycelia) is isolated from ganoderic acid and P. umbellatus and it exerts antiproliferative effects against malignant cells. However, no previous study has reported the inhibitory effects of Khz on the growth of human colon cancer cells. In the present study, we found that Khz suppressed cell division and induced apoptosis in HCT116 cells. Khz cytotoxicity was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Khz reduced cell viability and mitochondrial membrane potential levels and it also induced disruption of the mitochondrial membrane potential and increased calcium concentration and reactive oxygen species generation. Khz increased caspase 3, PARP, caspase 7, and caspase 9 levels, but reduced Bcl-2 protein levels. Flow cytometry showed that the percentage of HCT116 cells in the sub-G1 phase of the cell cycle increased in response to Khz treatment.

Khz-cp (crude polysaccharide extract obtained from the fusion of Ganoderma lucidum and Polyporus umbellatus mycelia) induces apoptosis by increasing intracellular calcium levels and activating P38 and NADPH oxidase-dependent generation of reactive oxygen species in SNU-1 cells.

BACKGROUND: Khz-cp is a crude polysaccharide extract that is obtained after nuclear fusion in Ganoderma lucidum and Polyporus umbellatus mycelia (Khz). It inhibits the growth of cancer cells. METHODS: Khz-cp was extracted by solvent extraction. The anti-proliferative activity of Khz-cp was confirmed by using Annexin-V/PI-flow cytometry analysis. Intracellular calcium increase and measurement of intracellular reactive oxygen species (ROS) were performed by using flow cytometry and inverted microscope. SNU-1 cells were treated with p38, Bcl-2 and Nox family siRNA. siRNA transfected cells was employed to investigate the expression of apoptotic, growth and survival genes in SNU-1 cells. Western blot analysis was performed to confirm the expression of the genes. RESULTS: In the present study, Khz-cp induced apoptosis preferentially in transformed cells and had only minimal effects on non-transformed cells. Furthermore, Khz-cp was found to induce apoptosis by increasing the intracellular Ca2+ concentration ([Ca2+]i) and activating P38 to generate reactive oxygen species (ROS) via NADPH oxidase and the mitochondria. Khz-cp-induced apoptosis was caspase dependent and occurred via a mitochondrial pathway. ROS generation by NADPH oxidase was critical for Khz-cp-induced apoptosis, and although mitochondrial ROS production was also required, it appeared to occur secondary to ROS generation by NADPH oxidase. Activation of NADPH oxidase was shown by the translocation of the regulatory subunits p47phox and p67phox to the cell membrane and was necessary for ROS generation by Khz-cp. Khz-cp triggered a rapid and sustained increase in [Ca2+]i that activated P38. P38 was considered to play a key role in the activation of NADPH oxidase because inhibition of its expression or activity abrogated membrane translocation of the p47phox and p67phox subunits and ROS generation. CONCLUSIONS: In summary, these data indicate that Khz-cp preferentially induces apoptosis in cancer cells and that the signaling mechanisms involve an increase in [Ca2+]i, P38 activation, and ROS generation via NADPH oxidase and mitochondria.

A new phenylethanoid glycoside with antioxidant and anti-HBV activity from Tarphochlamys affinis.

A new phenylethanoid glycoside, named taraffinisoside A (1), together with five known glycosides were isolated from the stems and leaves of Tarphochlamys affinis. The structure of taraffinisoside A was identified on the basis of detailed spectral analysis. Compounds 1-4 and 6 showed potent antioxidant activities with IC50 values of 10.36, 19.73, 43.95, 15.30 and 46.04 µM by 1,1-diphenyl-2-picryhydrazyl radical-scavenging assay. Compounds 1, 2 and 4 showed anti-HBV activities, with IC50 values of 0.50, 0.72 and 0.26 mM for HBsAg and 0.93, 0.42 and 0.07 mM for HBeAg, respectively.

Pressor mechanism evaluation for phytochemical compounds using in silico compound-protein interaction prediction.

In this study, a method was applied to evaluate pressor mechanisms through compound-protein interactions. Our method assumed that the compounds with different pressor mechanisms should bind to different target proteins, and thereby these mechanisms could be differentiated using compound-protein interactions. Twenty-six phytochemical components and 46 tested target proteins related to blood pressure (BP) elevation were collected. Then, in silico compound-protein interactions prediction probabilities were calculated using a random forest model, which have been implemented in a web server, and the credibility was judged using related literature and other methods. Further, a heat map was constructed, it clearly showed different prediction probabilities accompanied with hierarchical clustering analysis results. Followed by a compound-protein interaction network was depicted according to the results, we can see the connectivity layout of phytochemical components with different target proteins within the BP elevation network, which guided the hypothesis generation of poly-pharmacology. Lastly, principal components analysis (PCA) was carried out upon the prediction probabilities, and pressor targets could be divided into three large classes: neurotransmitter receptors, hormones receptors and monoamine oxidases. In addition, steroid glycosides seem to be close to the region of hormone receptors, and a weak difference existed between them. This work explored the possibility for pharmacological or toxicological mechanism classification using compound-protein interactions. Such approaches could also be used to deduce pharmacological or toxicological mechanisms for uncharacterized compounds.

Khz (fusion of Ganoderma lucidum and Polyporus umbellatus mycelia) induces apoptosis by increasing intracellular calcium levels and activating JNK and NADPH oxidase-dependent generation of reactive oxygen species.

Khz is a compound derived from the fusion of Ganoderma lucidum and Polyporus umbellatus mycelia that inhibits the growth of cancer cells. The results of the present study show that Khz induced apoptosis preferentially in transformed cells and had only minimal effects on non-transformed cells. Furthermore, Khz induced apoptosis by increasing the intracellular Ca(2+) concentration ([Ca(2+)](i)) and activating JNK to generate reactive oxygen species (ROS) via NADPH oxidase and the mitochondria. Khz-induced apoptosis was caspase-dependent and occurred via a mitochondrial pathway. ROS generation by NADPH oxidase was critical for Khz-induced apoptosis, and although mitochondrial ROS production was also required, it appeared to occur secondary to ROS generation by NADPH oxidase. Activation of NADPH oxidase was demonstrated by the translocation of regulatory subunits p47(phox) and p67(phox) to the cell membrane and was necessary for ROS generation by Khz. Khz triggered a rapid and sustained increase in [Ca(2+)](i), which activated JNK. JNK plays a key role in the activation of NADPH oxidase because inhibition of its expression or activity abrogated membrane translocation of the p47(phox) and p67(phox) subunits and ROS generation. In summary, these data indicate that Khz preferentially induces apoptosis in cancer cells, and the signaling mechanisms involve an increase in [Ca(2+)](i), JNK activation, and ROS generation via NADPH oxidase and mitochondria.

Development and validation of an LC-ESI-MS/MS method for the determination of nitidine chloride in rat plasma.

A new liquid chromatography-electrospray ionization-mass/mass spectrometry (LC-ESI-MS/MS) assay method has been developed and validated for the quantification of nitidine chloride (NC), an anti-cancer bioactive substance of Zanthoxylum nitidum (Roxb.) DC. plants, in rat plasma using carbamazepine as an internal standard (I.S.). The NC and I.S. were extracted from rat plasma by acetonitrile protein procedure. Chromatographic separation was carried out with a C(18) column (2.1 mm × 150 mm, 3 µm) with a security guard C18 column (4 mm × 20 mm, 3 µm). The mobile phase consisted of acetonitrile-10 mM ammonium acetate buffer solution-formic acid (35:65:0.2, v/v/v) and delivered at the flow rate of 0.25 mL/min. LC-ESI-MS/MS was performed on a triple-quadrupole mass spectrometry equipped with electrospray ionization (ESI) and positive multiple reaction monitoring (MRM). Target ions were monitored at [M](+)m/z 348.2 for NC and [M]⁺ m/z 237.2 for I.S. The method was linear over the concentration range of 5.0-1500.0 ng/mL. The intra- and inter-day relative standard deviations of the assay were less than 5.0%. The lower limit of quantification was 5.0 ng/mL. The developed method was successfully applied to the estimation of the pharmacokinetic parameters of NC by intravenous administration to rats.

Protective effect of Yulangsan polysaccharide on focal cerebral ischemia/reperfusion injury in rats and its underlying mechanism.

OBJECTIVE: To study the focal cerebral ischemia/reperfusion (I/R) injury induced by a middle cerebral artery occlusion (MCAO), and the effects of Yulangsan (YLS) polysaccharide on this injury. METHODS: This study took place in the Pharmacology Research Laboratory at Guangxi Medical University, China, between March and May 2007. Two hundred and forty rats were randomly divided into I/R group, sham-operated group, high-, medium-, and low-dose of YLS polysaccharide groups, and nimodipine (Nim) group. The animals were intragastrically administered with drugs for 7 days. An operation was performed to induce an MCAO model in the rats. Reperfusion was started after 2 hours of MCAO. The influences of YLS polysaccharide on the neurological score, the brain water content, the infarct volume, the activities of super oxide dismutase (SOD) and nitric oxide synthase (NOS), the contents of malondialdehyde (MDA) and nitric oxide (NO), the expressions of B-cell lymphoma/leukemia-2 (Bcl-2) and Bcl-2-associated X protein (Bax) in brain tissue were investigated; the morphological changes of rat cerebral cortical neurons were observed. RESULTS: Compared with the I/R group, YLS polysaccharide reduced the neurological score, the brain water content, the infract volume, MDA and NO contents, the NOS activity, and the expression of Bax, and increased SOD activity, and the expression of Bcl-2 in the brain tissue, and neuronal edema was reduced. CONCLUSION: The YLS polysaccharide has a protective effect on cerebral ischemia/ reperfusion injury; the mechanism may be related to attenuating free radicals, and increasing the Bcl-2/Bax ratio.

[Effect of cedemex on cAMP and cGMP levels of different brain areas in morphine withdrawal rats].

OBJECTIVE: To investigate the effect of Cedemex on cAMP and cGMP contents in different brain regions in morphine withdrawal rats precipitated by naloxone. METHOD: A physical morphine dependent model of rats was established by subcutaneous injection of morphine in gradually increasing dosage within 7 days. cAMP and cGMP contents of VTA, cortex and hippocampus of the rat brains were determined by radioimmunoassay. RESULT: The morphine withdrawal symptoms of rats were relieved significantly by ig Cedemex. Compared with the controls, cAMP content in the region of VTA, cortex and hippocampus of the morphine dependent rats were significantly higher (P < 0.05), while cGMP contents in those regions were significantly lower (P < 0.05). cAMP contents in the area of VTA, cortex and hippocampus of the morphine dependent rats were significantly reduced, while cGMP contents were significantly increased by ig Cedemex. CONCLUSION: Cedemex may significantly attenuate the morphine withdrawal symptoms in rats. The mechanism of this effect may be related to adjusting the contents of cAMP and cGMP in some brain regions.

[Effect of three flavones on enzyme activity of recombinant human phosphoinositide 3-kinase p110beta catalytic subunit].

OBJECTIVE: To study the effect of three flavones-luteolin, apigenin and genistein on activity of recombinant human phosphoinositide 3-kinase (PI3-K) p110beta catalytic subunit. METHODS: Recombinant human P13-K p110beta catalytic subunit was expressed by gene engineering. PI3-K activity was assayed by incubation recombinant PI3-K p110beta with phosphatidylinostiol-4,5-bisphosphate and [gamma-32P] ATP; the 32P-radiolabeled lipids were extracted with cholroform and methanol, and assessed by scintillation counter. RESULTS: Luteolin and apigenin showed inhibition on the recombinant p110beta catalytic subunit with IC50 8. 65 micromol/L and 11.56 micromol/L, but genistein had no inhibition. CONCLUSION: Luteolin and apigenin are inhibitors of P13-K. The recombinant P13-K p1100 catalytic subunit may be used as a molecular target for simpler screening and development of more effective inhibitors of P13-K.

[Observation of the promotion effect taurine on hepatic stellate cell's apoptosis in rat hepatic fibrosis model].

OBJECTIVE: To observe the effect of taurine on hepatic stellate cell's apoptosis induced by carbon tetrachloride (CCl4) in rats and to study its protective mechanisms. METHODS: CCl4-induced rat hepatic fibrosis was treated by taurine. Serum alanine aminotransferase (ALT), plasma protein, hyaluronic acid (HA), procollagen III (PC III), hepatic microsomal drug-metabolizing enzyme and anti-transforming growth factor beta1 (TGF-beta1) were determined. In addition, hepatic stellate cell's apoptosis and the pathological changes of liver tissue were observed under light microscope. RESULTS: The activity of serum ALT and the levels of serum HA, PC III were markedly reduced by taurine treatment. The hepatic cytochrome P450 (Cyt. P450) and cytochrome b5 (Cytb5) contents were increased by the same treatment. In addition, taurine could significantly inhibit the expression of TGF-beta1, promote the hepatic stellate cell's apoptosis, and relieve hepatic fibrosis. CONCLUSION: Taurine fulfills a role in promoting hepatic stellate cell's apoptosis in the case of hepatic fibrosis, it mitigates the liver injury, decreases the expression of TGF-beta1, and relieves hepatic fibrosis.