Objective To observe the effect of muscone on the blood-brain barrier and neuropathological changes after complete cerebral ischemia/reperfusion of rats.Methods The global cerebral ischemia/reperfusion injury model was established by the four-vessel occlusion(4VO) method,and then the model animals were given high-,middle-and low-dose muscone.Meanwhile,sham-operation group,model group,and positive control group were also set up.The rats neuroethology behavior,and the contents of superoxide dismutase(SOD),malondialdehyde(MDA) and excitatory amino acid(EAA) in ischemic brain tissue were examined.Results Muscone increased SOD content,lowered MDA content,reduced the increase of EAA content caused by ischemia and hypoxia,and inhibited the excitatory neurotoxicity caused by EAA.Conclusion Muscone has obvious protective effect on rats with complete cerebral ischemia/reperfusion injury.

Objective To develop an HPLC method for the determination of chlorogenic acid in Xiao'er Yinniu Granula.Methods A Phenomenex ODS C18 column (4.6 mm?250 mm,5 ?m) was used,column temperature was at 20 ℃,methanol-0.4 %phosphoric acid water solution(20:80)was used as the mobile phase,the flow rate was 1 mL?min-1,and the detection wavelength was 327 nm.Results The calibration curve of chlorogenic acid was linear between 0.284～1.704 ?g (r=0.999 6),the average recovery and the relative standard deviation were 96.19 %and 1.16 %(n=5),respectively.Conclusion The method is simple,accurate and reproducible,by whihc chlorogenic acid can be better separated from Xiao'er Yinniu Granula.This method can be used for quality control of Xiao'er Yinniu Granula.

OBJECTIVETo study the time-toxicity and dose-toxicity relationships caused by multiple dose of total Bupleurum saponin extracts to rats.

METHODRats were picked according to different time or dose points, and total Bupleurum saponin crude extracts were administered to rats. The death circumstance and toxicity of mice were observed, ALT and AST in serum were detected. Indice of the liver, index was calculated. And the morphological changes of liver tissue were observed under light microscope.

RESULTThe "time-toxicity" relationship study showed that ALT and AST in rats' serum began to increase after 7 days' administration, and with obvious hepatotoxicity after 15 days', then there was concurrent toxicity and death. The "dose-toxicity" relationship study showed that compared with the control group, the total Bupleurum saponin crude extracts between 51. 2 and 125.0 g x kg(-1) dose could cause the obvious hepatotoxicity to rats in 15 days' administration, which was represented by that ALT and AST in serum increased significantly with the dose increased, the ratio of liver to body increased, and under light microscope, the different doses' liver tissue of mice all had edema in different degree and fatty degeneration in liver cells, and the high-dose and long time administration group appeared to be necrosis, lobular structure unclear. The above-mentioned change gradually aggravated with dose increasing, and obviously diversity compared with distilled water control group.

CONCLUSIONWith administrated a multiple dose and long time to rats, the total Bupleurum saponins crude extracts could cause serious liver injury and even death, and there were certain time-toxicity and dose-toxicity relationships.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Bupleurum ; chemistry ; Complex Mixtures ; administration & dosage ; toxicity ; Dose-Response Relationship, Drug ; Drug Administration Schedule ; Female ; Liver ; drug effects ; enzymology ; pathology ; Male ; Rats ; Rats, Wistar ; Saponins ; chemistry ; Time Factors ; Toxicity Tests ; methods

Objective To explore the molecular mechanisms by which mitochondrial estrogen receptorβ(ERβ) suppresses non-small cell lung cancer cell apoptosis induced by apoptotic stimulations. Methods The mitochondrial localization of ERβin non-small cell lung cancer cell lines A549 and 201T was determined using immunofluorescence and Western blotting. The changes of apoptosis of the cells with mitochondrial ERβ overexpression or knockdown in response to cisplatin and STS treatments were assessed, and mitochondrial ERβ interaction with the pro- apoptotic protein Bad was detected using co-immunoprecipitation and Western blotting. Results ERβ was localized in the mitochondria in A549 and 201T cells. ERβoverexpression significantly reduced while ERβknockdown increased Bax activation and cell apoptosis induced by cisplatin and STS. Mitochondrial ERβinteraction with pro-apoptotic protein Bad may suppress Bax activation and its translocation to the mitochondria. Conclusion Mitochondrial ERβ can suppress apoptosis of non-small cell lung cancer cells induced by cisplatin or STS through interaction with Bad, suggesting the value of mitochondrial ERβ as a new therapeutic target for treatment of non-small cell lung cancer.

Objective To explore the molecular mechanisms by which mitochondrial estrogen receptorβ(ERβ) suppresses non-small cell lung cancer cell apoptosis induced by apoptotic stimulations. Methods The mitochondrial localization of ERβin non-small cell lung cancer cell lines A549 and 201T was determined using immunofluorescence and Western blotting. The changes of apoptosis of the cells with mitochondrial ERβ overexpression or knockdown in response to cisplatin and STS treatments were assessed, and mitochondrial ERβ interaction with the pro- apoptotic protein Bad was detected using co-immunoprecipitation and Western blotting. Results ERβ was localized in the mitochondria in A549 and 201T cells. ERβoverexpression significantly reduced while ERβknockdown increased Bax activation and cell apoptosis induced by cisplatin and STS. Mitochondrial ERβinteraction with pro-apoptotic protein Bad may suppress Bax activation and its translocation to the mitochondria. Conclusion Mitochondrial ERβ can suppress apoptosis of non-small cell lung cancer cells induced by cisplatin or STS through interaction with Bad, suggesting the value of mitochondrial ERβ as a new therapeutic target for treatment of non-small cell lung cancer.

Objective To investigate the correlation between miR-221 and epithelial-mesenchymal transition (EMT) in drug-resistant glioma cells. Methods The expression levels of miR-221, PTEN, p-Akt, E-cadherin, vimentin, and MRP1 were quantitatively analyzed in Z1 cells (primary drug-resistant cells), Z2 cells (drug-sensitive cells) and Z2-BCNU cells (drug-resistant cells) using fluorescent real-time PCR and Western blotting. Results The expression levels of PTEN were significantly increased in Z2 cells compared with Z1 and Z2-BCNU cells which overexpressed miR-221 and vimentin. The expression levels of vimentin, p-Akt and MRP1 were significantly decreased in Z2 cells overexpressing E-cadherin. Conclusion MiR-221 regulates the expression of EMT-related genes through down-regulation of PTEN and activation of PI3-K/Akt signaling.

Objective To investigate the correlation between miR-221 and epithelial-mesenchymal transition (EMT) in drug-resistant glioma cells. Methods The expression levels of miR-221, PTEN, p-Akt, E-cadherin, vimentin, and MRP1 were quantitatively analyzed in Z1 cells (primary drug-resistant cells), Z2 cells (drug-sensitive cells) and Z2-BCNU cells (drug-resistant cells) using fluorescent real-time PCR and Western blotting. Results The expression levels of PTEN were significantly increased in Z2 cells compared with Z1 and Z2-BCNU cells which overexpressed miR-221 and vimentin. The expression levels of vimentin, p-Akt and MRP1 were significantly decreased in Z2 cells overexpressing E-cadherin. Conclusion MiR-221 regulates the expression of EMT-related genes through down-regulation of PTEN and activation of PI3-K/Akt signaling.

OBJECTIVETo explore the effect of CD99 overexpression on the morphology and differentiation-related phenotypes of classical Hodgkin's lymphoma cell line L428 and investigate the role of CD99 gene in Hodgkin/Reed-Sternberg (HRS) cell generation and transformation.

METHODSThe effect of CD99 overexpression on the cell morphology was detected by HE staining and phalloidin staining. Differentiation-related protein expressions were detected by immunocytochemistry and flow cytometry after stable transfection of CD99 gene in L428 cells.

RESULTSCD99 overexpression caused a decrease of the cell size and reorganization of the actin cytoskeleton in L428 cells. Upregulation of CD99 led to the loss of classical Hodgkin's lymphoma diagnosis marker CD30 and CD15 and the restoration of the B-cell makers of PAX5, CD19, CD79α, BCL-6, and CD10.

CONCLUSIONCD99 overexpression leads to redifferentiation of L428 cells towards B cells, suggesting that the loss of B-cell phenotype in classical Hodgkin's lymphoma is very likely a result of down-regulated CD99 expression.

12E7 Antigen ; Antigens, CD ; genetics ; B-Lymphocytes ; cytology ; Cell Adhesion Molecules ; genetics ; Cell Differentiation ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Hodgkin Disease ; genetics ; pathology ; Humans

OBJECTIVETo explore the molecular mechanisms by which mitochondrial estrogen receptor β (ERβ) suppresses non-small cell lung cancer cell apoptosis induced by apoptotic stimulations.

METHODSThe mitochondrial localization of ERβ in non-small cell lung cancer cell lines A549 and 201T was determined using immunofluorescence and Western blotting. The changes of apoptosis of the cells with mitochondrial ERβ overexpression or knockdown in response to cisplatin and STS treatments were assessed, and mitochondrial ERβ interaction with the pro-apoptotic protein Bad was detected using co-immunoprecipitation and Western blotting.

RESULTSERβ was localized in the mitochondria in A549 and 201T cells. ERβ overexpression significantly reduced while ERβ knockdown increased Bax activation and cell apoptosis induced by cisplatin and STS. Mitochondrial ERβ interaction with pro-apoptotic protein Bad may suppress Bax activation and its translocation to the mitochondria.

CONCLUSIONMitochondrial ERβ can suppress apoptosis of non-small cell lung cancer cells induced by cisplatin or STS through interaction with Bad, suggesting the value of mitochondrial ERβ as a new therapeutic target for treatment of non-small cell lung cancer.

Apoptosis ; Carcinoma, Non-Small-Cell Lung ; pathology ; Cell Line, Tumor ; Cisplatin ; Estrogen Receptor beta ; metabolism ; Humans ; Mitochondrial Proteins ; metabolism ; bcl-Associated Death Protein ; metabolism

OBJECTIVETo explore the mechanism of glutamate (Glu)-induced PC12 cell apoptosis and the protective effect of muscone.

METHODPC12 cells were randomly divided into six groups: normal group (Normal), model group injured by glutamate (Glu), nimodipine group (Nim) and muskone groups (Mus) of high, middle and low doses. The PC12 cells were pretreated with or without different concentrations of muskone for 30 min and then exposed to glutamate. MTT assay for cell survival, flow cytometric detection of apoptotic cells, DCF assay for reactive oxygen species (ROS) and flow cytometric assay was performed to determine the mitochondrial membrane potential in PC12 cell.

RESULTPC12 cell damage, the concentrations of [Ca2+], and apoptosis induced by Glu were decreased after being administrated with paeoniflorin.

CONCLUSIONMuskone inhibited Glu-induced apoptosis in PC12 cells. The mechanism is related to inhibiting intracellular Ca2+ overload and maintaining mitochondral membrance potential.

Animals ; Apoptosis ; drug effects ; Calcium ; metabolism ; Cell Survival ; drug effects ; Cycloparaffins ; pharmacology ; Glutamic Acid ; pharmacology ; Membrane Potential, Mitochondrial ; drug effects ; PC12 Cells ; Pheochromocytoma ; metabolism ; physiopathology ; Protective Agents ; pharmacology ; Rats