Adenocarcinoma, Clear Cell ; complications ; metabolism ; pathology ; surgery ; Adenoma ; pathology ; Adnexal Diseases ; pathology ; Adult ; Carcinoma, Renal Cell ; pathology ; secondary ; Cystadenoma, Papillary ; complications ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Fallopian Tube Neoplasms ; complications ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Keratin-7 ; metabolism ; Kidney Neoplasms ; pathology ; secondary ; Mucin-1 ; metabolism ; von Hippel-Lindau Disease ; complications ; metabolism ; pathology ; surgery

OBJECTIVE: To observe the effect of rosiglitazone on the production of nitric oxide (NO) and the expression of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB) /the endothelial nitric oxide synthase (eNOS) in cultured human umbilical vein endothelial cells(HUVECs), and to investigate the mechanism of signal transduction of rosiglitazone in improving the endothelial function. METHODS: HUVECs were treated with various concentrations of rosiglitazone. The NO level was measured using Griess Reaction in cell culture supernatants; the expressions of PI3K-, PKB- and eNOS mRNA were measured using RT-PCR; and the expressions of PKB, eNOS, and phosphorylation of PKB-Ser473, eNOS-Ser1177 were measured using Western Blot. RESULTS: Rosiglitazone increased the endothelial NO production in a dose- and time-dependent manner in cultured HUVECs, and also increased the expression of PI3K mRNA and the phosphorylation of PKB-Ser473 and eNOS-Ser1177 in a concentration-dependent manner, with no alteration in the expression of PKB and eNOS in cultured HUVECs. N(w)-nitro-L- arginine methyl ester (L-NAME, eNOS synthase inhibitor) blocked the rosiglitazone-induced NO formation; LY294002 (a PI3K inhibitor) prevented the NO production; and the phosphorylation of eNOS and PKB was induced by rosiglitazone. CONCLUSION Treatment with rosiglitazone can increase the NO production and improve the endothelial function through up-regulating the PI3K/PKB/eNOS signal pathways in cultured HUVECs.
Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; drug effects ; Proto-Oncogene Proteins c-akt ; metabolism ; Rosiglitazone ; Signal Transduction ; drug effects ; Thiazolidinediones ; pharmacology

0.05).Conclusions SNP of 2350G→A in ACE gene is associated with MI,AA genotype is probably a genetic marker of MI in Han nationality.

OBJECTIVETo observe the changes of aortic endothelium-dependent vasodilation function (EDVR) and expressions of endothelial nitric oxide synthase (eNOS), phosphatidylinositol 3-kinase (PI3K), and protein kinase B (PKB) in insulin-resistance (IR) and type 2 diabetic rats.

METHODSIR rat model was established by feeding 4-6 week-old male SD rats with high glucose and cholesterol diet for 6 weeks and type 2 diabetes (DM) were induced by intraperitoneal injection with low dose streptozotocin (STZ) to IR rats. Glucose infusion rate (GIR) was determined by euglycemic hyperinsulinemic clamp technique, EDVR by acetylcholine (Ach)-induced vasodilation response in isolated aortic rings, aortic NO concentration by Griess Reaction, activation of eNOS detected by immunohistochemical SP method, mRNA expressions of eNOS-, PI3K- and PKB of aorta were assayed by RT-PCR, aorta ultrastructure observed by electron microscopy. Body weight, fast plasma glucose (FPG), insulin (FINS), triglyceride (TG), cholesterol (TC) were determined and the insulin sensitivity index (ISI) was calculated.

RESULTS(1) Body weight, FINS, TG and TC levels were significantly higher while ISI and GIR significantly lower in IR and DM rats than that in normal control rats (P < 0.05). (2) Aorta EDVR decreased significantly in IR and DM group compared with that in control group (P < 0.05) and EDVR was also significantly reduced in DM rats than that in IR rats (P < 0.05). The maximum Ach-induced vasodilation response (EDVR(max), P < 0.01) was positively correlated with ISI and negatively correlated with FPG, TG, TC and FINS (P < 0.01). (3) Aortic NO concentration, the mRNA expressions of eNOS-, PI3K-, and PKB and eNOS immunohistochemical expression in aorta were significantly lower in IR and DM rats compared with normal control rats and the decrease was more pronounced in DM rats (P < 0.05 vs. IR). (4) Pathologic aortic ultrastructure changes were also visualized in IR and DM rats.

CONCLUSIONOur results suggest that reduced NO concentration and expression as well as reduced PI3K-, PKB-, and eNOS mRNA expressions might contributed to the reduced EDVR function and related pathological ultrastructure changes in IR and DM rats.

Animals ; Aorta ; metabolism ; physiopathology ; Diabetes Mellitus, Experimental ; metabolism ; physiopathology ; Diabetes Mellitus, Type 2 ; metabolism ; physiopathology ; Endothelium, Vascular ; metabolism ; physiopathology ; Insulin Resistance ; Male ; Nitric Oxide Synthase Type III ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Rats, Sprague-Dawley ; Vasodilation

OBJECTIVE: To investigate the change of carotid intima-media thickness (IMTc) and serum matrix metalloproteinases-9 (MMP-9) levels in newly diagnosed Type 2 diabetic patients, and to analyze the relationship between MMP-9 and IMTc; at the same time, to assess the effect of rosiglitazone on IMTc and MMP-9 levels. METHODS: Fifty-eight patients with Type 2 diabetes mellitus were selected in our study, and 25 healthy adults served as normal controls. Diabetic patients were divided into 2 groups: Group A (31 subjects) were treated with rosiglitazone (4 mg/d), and Group B (27 subjects) were treated with metformin alone (500 approximately 1,500 mg/d). They all received the treatment for 3 months. The IMTc was measured by high resolution ultrasonography, and the serum MMP-9 was determined by enzyme linked immunosorbent assay (ELISA) to assess the relationship between IMTc and MMP-9. RESULTS: The mean level of serum IMTc and MMP-9 in Type 2 diabetic patients was significantly higher than that in healthy adults (P < 0.05). After treatment with rosiglitazone and metformin, IMTc and serum MMP-9 levels decreased significantly (P < 0.05). There was no obvious change in IMTc and serum MMP-9 levels in group B before and after the treatment (P = 0. 071, P = 0.065). Using multiple linear stepwise regression analysis, the significant correlation between IMTc and HbA1C, BMI, WHR, HDL-C, MMP-9 were discovered. CONCLUSION IMTc and MMP-9 levels increase in newly diagnosed Type 2 diabetic patients, suggesting that there is closely relationship between serume MMP-9 levels and early diabetic macrovascular disease. IMTc and MMP-9 can be reduced significantly in the newly diagnosed diabetic patients after being treated with rosiglitazone, which may be one of the protective mechanisms of vascular vessels.
Adult ; Carotid Arteries ; pathology ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; pathology ; Female ; Humans ; Hypoglycemic Agents ; therapeutic use ; Male ; Matrix Metalloproteinase 9 ; blood ; Middle Aged ; Rosiglitazone ; Thiazolidinediones ; therapeutic use ; Tunica Intima ; pathology ; Tunica Media ; pathology

OBJECTIVETo observe the effect of Tanshinone II A on the expression of epidermal growth facter (EGF) and epidermal growth facter recepter (EGFR) in human hepatocellular carcinoma cell line SMMC-7721.

METHODSThe human hepatocellular carcinoma SMMC-7721 cells cultured in vitro was treated with different concentrations of Tanshinone II A. The proliferation of the cells was measured by MTT assay, and the apoptosis of the cells was investigated by flow cytometry and cytochemical staining with Hoechst 33342. The expression of EGF and EGFR was detected by immunocytochemistry method. The levels of EGF in medium were measured by radioimmunoassay.

RESULTTanshinone II A inhibited the growth of SMMC-7721 cells remarkably in a dose-dependent manner. The inhibitory rate reached the peak (72.5%) after 0.5 microg/ml Tanshinone II A was used for 48 h, which was significantly higher than that in the controls (P<0.05). FCM analysis showed that when SMMC-7721 cells were treated with 0.5 microg/ml Tanshinone II A, the apoptosis rates for 24 h, 48 h and 72 h were (4.06+/-0.27)%, (7.58+/-0.56)% and (5.23+/-0.13)%, respectively which were markedly higher than those in the controls (all P<0.01). SMMC-7721 cell apoptosis with cell shrinkage, nuclear chromatin concentration and fragmentation as well as the formation of apoptotic bodies were observed by cytochemical staining when treated with Tanshinone II A. The immunocytochemistry showed that the expressions of EGF and EGFR were down regulated while the concentration of Tanshinone II A was increasing. The high expression rates for EGF and EGFR were 10%, 20%, respectively, and the gray scale was 181.52+/-1.63, 179.37+/-1.59, which were markedly higher than those in the controls (all P<0.05). The levels of EGF in medium measured by radioimmunoassay were decreased significantly after Tanshinone II A treatment.

CONCLUSIONTanshinone II A can inhibit cell proliferation and induce apoptosis in hepatocellular carcinoma cell line SMMC-7721, which may be related to the down-regulation of EGF and EGFR protein expression.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Diterpenes, Abietane ; Down-Regulation ; drug effects ; Epidermal Growth Factor ; genetics ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Phenanthrenes ; pharmacology ; Receptor, Epidermal Growth Factor ; genetics ; metabolism

Acupuncture Therapy ; methods ; Adult ; Female ; Humans ; Male ; Middle Aged ; Nerve Compression Syndromes ; therapy ; Skin ; innervation ; Upper Extremity ; innervation

OBJECTIVETo study the effects of in vitro inducement on the expression of SF1-G imprinted genes, Kcnq1 and Cdkn1c during the course of differentiation from mouse embryonic stem (ES) cells to islet-like cells.

METHODSMouse ES cells were induced to differentiate into islet-like cells in vitro. The expression of islet specific markers was tested by RT-PCR or immunofluorescence. RT-PCR/RFLP was used to test the imprinted genes parental expression in cells at different stages.

RESULTSIslet specific genes, such as Insulin, Glucagon, Somatostatin, IAPP and Glut2, were expressed in differentiated cells. The proteins of insulin, C-peptide and Somastatin were expressed in the final stage cells. Imprinted gene Kcnq1 and Cdkn1c were biallelicly expressed in islet-like cells.

CONCLUSIONSMouse ES cells can be successfully induced into islet-like cells in vitro. Gene imprinting status of Kcnq1 and Cdkn1c may be changed in differentiated cells (causing loss of imprinting) during the in vitro inducement.

Animals ; Cell Differentiation ; drug effects ; Insulin ; Islets of Langerhans ; cytology ; Mice ; Mouse Embryonic Stem Cells ; Proteins ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells

Child, Preschool ; Diagnosis, Differential ; Female ; Humans ; Rickets ; diagnosis ; etiology ; therapy ; Treatment Outcome ; Vitamin D Deficiency ; complications

OBJECTIVETo observe the relaxant effect of Aike Mixture (AKM) on isolated bladder and prostatic urethral smooth muscle of rabbits.

METHODSThe isolated bladder and prostatic urethral smooth muscle from male rabbits were placed in a Magnus bath and smooth muscle contraction was measured using a biological signal acquisition and analysis system. The effects of AKM in combination with methoxyamine, carbachol and CaCl2 on the contractile tension of muscle strips were determined by cumulative dosing.

RESULTSAKM dose-dependently reduced contractile tension of bladder trigone smooth muscle (r=0.831, P<0.05), reduced contractile wave amplitude (r=0.837, P<0.05) and decreased contractile frequency (r=-0.917, P<0.01). AKM significantly inhibited the increases in smooth muscle contraction induced by methoxyamine, carbachol and CaCl2.

CONCLUSIONAKM dose-dependently inhibited the contraction of rabbit isolated bladder and prostatic urethral smooth muscle by antagonizing α1-adrenergic receptors and M-cholinergic receptors.

Animals ; Calcium Chloride ; pharmacology ; Carbachol ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Hydroxylamines ; pharmacology ; In Vitro Techniques ; Male ; Muscle Contraction ; drug effects ; Muscle, Smooth ; drug effects ; physiology ; Neuromuscular Agents ; pharmacology ; Prostate ; drug effects ; physiology ; Rabbits ; Receptors, Adrenergic, alpha-1 ; metabolism ; Receptors, Muscarinic ; metabolism ; Urethra ; drug effects ; physiology ; Urinary Bladder ; drug effects ; physiology