Objective: To investigate the apoptosis inducing effects of cryptotanshinone (CPT) in human chronic myeloid leukemia K562 cells and its molecular mechanisms. Methods: K562 cells were firstly treated with CPT at different concentration. Then, the inhibitory effect of CPT was detected by MTT assay; morphological changes of cells were observed by inverted microscope; cell apoptosis was detected by Annexin V-FITC/PI staining. Western blotting was carried out to determine the expression of apoptosis-related proteins, including Caspase-3, Caspase-9, PARP, Bax, Bcl-2 and cytochrome C (Cyt C). The mitochondrial membrane potential was measured using JC-1 probe. Finally, the effects of CPT on the proliferation and apoptosis were detected in the absence or presence of inhibitors for mitochondrial permeability transition pore (PTP) and caspase-family. Results: CPT significantly inhibited the proliferation, induced apoptosis and morphological changes of K562 cells. Furthermore, CPT increased the Bax/Bcl-2 ratio obviously, decreased the mitochondrial membrane potential, and enhanced the Cyt C release. CPT also significantly increased the activities of pro-apoptotic proteins, such as Caspase-3, Caspase-9, and PARP. The inhibitors of PTP and caspase-family reduced the pharmacodynamics of CPT obviously. Conclusion: These results show that CPT could significantly induce the apoptosis of human chronic myeloid leukemia cells, in which its function is related with the mitochondrial pathway.

It is well documented that altered biosynthesis of cell surface N-linked oligosaccharides is associated with the transformed cells and tumors.N-acetylglucosaminyltransferase Ⅱ(GnT-II;EC 2.4.1.143)is a medial Golgi enzyme that catalyses the incorporation of a GlcNAc residue in ?-1,2 linkage to the Man-?-1,6 arm of the N-glycan core.This is an essential step in the biosynthetic pathway leading from hybrid to complex N-glycans.Because functional GnT-Ⅱ is an prerequisite of N-Acetylglucosaminyltransferase V performance,It was speculated that GnT-Ⅱ was involved in cancer development and progression.The expression of GnT-Ⅱ in mouse breast cancer cells 67NR and 4T1 which have different behavior of metastasis was analysed using RT-PCR.The amounts of GnT-Ⅱ in the highly metastatic cell 4T1 increased to 1.53 times of the lowly metastatic cell 67NR.To determine the association of GnT-Ⅱ with tumor progression,the GnT-Ⅱ encoding gene was amplified with RT-PCR and cloned into retrovirus vector pMSCV,resulting in pMSCV-GnT-Ⅱ.The recombinant plasmid was transfected into 4T1 and the transfected cells were selected in the medium containing puromycin,which were harvested to detect the adhesion ability to fibronection and the migration potential by transwell system.The cell adhesion to fibronectin was weakened by 67% and migration potential was increased by 82%.The data indicates that GnT-Ⅱ mediates cell adhesion and migration,thus may play an important role in cancer metastasis.

Objective: To study the effects of dihydrotanshinone (DHT) on the migration and invasion of human gastric cancer SGC7901 cells and to investigate its mechanism. Methods: SGC7901 cells were treated with different concentration of DHT. Then, the inhibitory effect of DHT was detected by MTT assay. The scratch adhesion test and Transwell assay were performed to determine the migration and invasion capacity of the cells. Quantitative PCR and Western boltting were used to examine the expression of MMP2, MMP9, Gli1, and HHIP in SGC7901 cells. Results: DHT could inhibit the proliferation of SGC7901 cells with obvious dose- and time- dependent effects. DHT significantly inhibited the migration and invasion ability in SGC7901 cells in vitro. The expression of MMP9 was obviously down-regulated after DHT treatment. Furthermore, DHT significantly inhibited the Gli1 mRNA and proteins expression levels, and evaluated the expression of HHIP in SGC7901 cells. Conclusion: DHT could inhibit the capability of migration and invasion of human gastric cancer SGC7901 cells. The potential molecular mechanism may be related to the inhibition of MMP9, and down-regulation of Hedgehog signaling pathway.

Actins ; metabolism ; Animals ; Collagen Type III ; metabolism ; Collagen Type IV ; metabolism ; Desmin ; metabolism ; Diabetes Mellitus, Experimental ; complications ; Diabetic Nephropathies ; metabolism ; pathology ; HSP47 Heat-Shock Proteins ; metabolism ; Kidney Glomerulus ; metabolism ; pathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Vimentin ; metabolism

Cryptotanshinone (CPT), a lipid soluble active compound in Salvia miltiorrhiza, has a significant inhibitory effect on multiple malignant tumors, e. g. chronic myeloid leukemia (CML) cells and can effectively enhance imatinib's chemotherapeutic effect. However, its functional molecular mechanism remained unclear. In this experiment, the authors conducted a systematic study on the effect of CPT on the imatinib sensitivity and P-glycoprotein (P-gp) expression in CML cells by using CML cells K562 and imatinib persister K562-R. The MTT assays were performed to determine CPT's impact on the inhibitory effect of imatinib. Annexin V-FITC/PI staining analysis was used to detect the changes in the cell apoptosis rate. The active changes in apoptosis regulatory proteins Caspase-3, Caspase-9 and PARP were determined by Western blot. After the cells were pretreated with the gradient concentration of CPT, the expression of P-gp was analyzed by Western blot and flow cytometry. The changes in intracellular concentrations of imatinib were determined by HPLC analysis. The results indicated that the pretreatment with CPT significantly increased the proliferation inhibiting and apoptosis inducing effects of imatinib on K562 and K562-R cells as well as the degradation product expression of pro-apoptotic proteins Caspase-3, Caspase-9 and PARP, with a significant difference with the control group (P < 0.01). However, CPT showed no impact on the P-gp expression in CML cells and the intracellular concentrations of imatinib. In summary, the findings suggested that CPT enhanced the sensitivity of CML cells to imatinib. Its mechanism is not dependent on the inhibition in P-gp expression and the increase in intracellular drug concentration.
ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Caspase 9 ; genetics ; metabolism ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Imatinib Mesylate ; pharmacology ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; genetics ; metabolism ; physiopathology ; Phenanthrenes ; pharmacology

Objective To investigate the mRNA and protein expressions of Nav 1.1 and Nav 1.2 in hippocampus following traumatic brain injury ( TBI) in rats.Methods After the lateral fluid percussion model was established in adult male Sprague Dawley rats,the rats were sacrificed at 2,12,24 and 72 hours after percussion and collected ipsilateral hippocampus for detecting mRNA and protein expressions of Nav 1.1 and Nav 1.2 by means of fluorescent quantitation RT-PCR,Western blot and immunofluo rescence staining.Results The mRNA expressions of Nav 1.1 and Nav 1.2 were significantly down-regulated (P<0.01) in hippocampus and reached the lowest level at 2 hours following TBI.The protein expression of Nav 1.1 was significantly down-regulated (P<0.01) but recovered near to level of control group at 72 hours after TBI.While there was no statistical difference on protein expression of Nav 1.2 in hippocampus after TBI compared with control group (P>0.05).Conclusion TBI induces significant down-regulated mRNA and protein expressions of Nav 1.1 in the hippocampus,which may be one of molecular mechanisms for functional alternation of sodium channels and excitotoxic action following TBI.

Objective To explore the impact of metabolic syndrome (MS)in bone mineral density (BMD) in elderly male type 2 diabetics (T2DM). Methods One hundred and fourty cases were divided into MS group and non-MS(NMS)group according to with or without MS. Height,weight were measured, body mass index(BMI) were calculated and the total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), glycated hemoglobin (HbA1c)were tested. The bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry (DEXA). The differences among these groups were compared and the risk factors for MS were analyzed. Results The MD, BMI, TC,TG in MS group was higher than that in NMS group (P < 0.05 vs. P < 0.01),and HDL-C was lower than NMS group (P < 0.05). Correlation analysis showed that BMD was negatively correlated with age , TG , HDL-C , and was positively correlated with BMI.The results of multiple stepwise regression analysis showed that age ,HDL-C,BMI were independent risk factors for BMD. Conclusion The BMD significantly increases in T2DM with MS.Obesity and low HDL-C have positive effects in bone mass.

Objective To report the clinical effect of primary percutaneous coronary intervention(PCI)in patients with acute myocardial infarction(AMI).Methods A retrospective study was accomplished on the clinical data of 13 AMI patients who underwent PCI from March 2004 to April 2006.Results The infarct-related artery (IRA)was successfully recanalized by primary PCI for 12 AMI patients,without major complications occurred in these cases during hospitalization.Conclusion Primary PCI should be firstly chosen for treatment of AMI in the hospitals which could carry out PCI.

OBJECTIVETo develop a method for the determination of salvianolic acid B and paeoniflorin in Shuangdan granules.

METHODThe chromatographic separation was performed on a Zorbax SB C18 column with a linear gradient elution of phosphoric acid-water (0.05:100) and phosphoric acid-acetonitrile-methanol (0.05:4:96) was used. The flow rate was 1 mL x min(-1) and column temperature was set at 25 degrees C. The UV detection wavelength was set at 230 nm.

RESULTThe linear ranger was of 0.710-22.7 microg for salvianolic acid B and 0.103-3.30 microg for paeoniflorin. Three batches of Shuangdan granules were analyzed, and the content of salvianolic acid B ranged from 6.46-11.6 mg x g(-1) and paeoniflorin ranged from 1.44-1.78 mg x g(-1).

CONCLUSIONThe method was accurate, reproducible, reliable, and could be used for quantitative control of Shuangdan granules.

Benzoates ; analysis ; Benzofurans ; analysis ; Bridged-Ring Compounds ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Glucosides ; analysis ; Monoterpenes ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo study whether anxiety and depression are associated with the development of neurogenic urination in children.

METHODSA total of 136 9 to 12-year-old children with neurogenic urination (case group) and 136 age-matched healthy children (control group) were enrolled. The Screen for Children Anxiety Related Emotion Disorders (SCARED) and Depression Self-rating Scale for Children (DSRSC) were used to evaluate the psychological status. The incidences of anxiety and depression as well as the SCARED and DSRSC scores were compared between two groups. Logistic regression analysis model was used to evaluate the relationship between psychological status and the development of neurogenic urination.

RESULTSThe case group was found to have a higher incidence of anxiety and depression compared with the control group (P<0.01). The SCARED score in the case group (28.1 ± 8.6) increased significantly compared with 14.4 ± 4.9 in the control group (P<0.01). The DSRSC score in the case group was also significantly higher than in the control group (13.5 ± 4.8 vs 9.1 ± 3.2; P<0.01). The logistic regression analysis showed that the children with anxiety (SCARED-score>23) had a 1.224-fold increased risk for the development of neurogenic urination compared with the children with the SCARED-score≤23 and that the children with depression (DSRSC-score&ge;15) had a 1.148-fold increased risk for the development of this disorder.

CONCLUSIONSAnxiety and depression participate in the development of neurogenic urination in school children.

Anxiety ; complications ; Child ; Depression ; complications ; Female ; Humans ; Logistic Models ; Male ; Urination Disorders ; etiology ; psychology