Objective To investigate the effects of xuanzhi analgesic tablets on plasma levels of 6-keto-prostaglandin Flα(6-K-PGFlα) and thromboxane B2(TXB2) in rats with acute blood stasis. Methods Sixty SD rats were divided into six groups randomly, namely normal control, model control, positive control, xuanzhi analgesic tablets at 1. 36, 2. 72, and 5. 44 g·kg-1 groups. The rat model of blood stasis syndrome was caused by subcutaneous injection of adrenaline incorporated with ice-bathing. The effects of xuanzhi analgesic tablet on 6-K-PGFlαand TXB2 were observed. Results Compared with the normal control,plasma level of 6-K-PGFlα was significantly reduced(P<0. 01) and that of TXB2 in the model control was evidently increased(P<0. 01). Three dosages of xuanzhi analgesic tablets significantly raised 6-K-PGFlαlevel(P<0. 05)and lowered TXB2 level (P<0. 05). Conclusion Xuanzhi analgesic tablets significantly adjust plasma levels of 6-K-PGFlαand TXB2in rats with acute blood stasis. Xuanzhi analgesic tablets can coreect the imbalance between PGI2 and TXA2 through increasing 6-K-PGFlαand desearing TXB2 levels.

Aim To establish insulin resistance cell model on HepG2 cells (human embryonic liver tumor cells )and investigate the effect of berberine hydro-chloride on insulin-resistant HepG2 (IR-HepG2 ) cells.Methods ① IR model was induced by respec-tively using 10 -9 ,10 -8 ,10 -7 ,10 -6 ,10 -5 ,10 -4 mol ·L-1 insulin with 25 mmol · L-1 glucose in HepG2 cells.② HepG2 cells were incubated with 2-NBDG (fluorescent labeled glucose)in a series of concentra-tion:50,100,200,400,600,800 μmol·L-1 and a series of incubation time:20,40,60,80,100 min, to select the optimum concentration of insulin and the optimum incubation concentration and time of 2-NBDG in HepG2 cells.The success of the model was deter-mined by detecting the consumption of glucose in the cell supernatant and the uptake of glucose in HepG2 cells.③To study the effect of berberine hydrochloride on improving insulin resistance on the cell level,met-formin and berberine hydrochloride were used in the IR cells.Results Six concentrations of insulin induced the IR model in different degrees.Although 10 -4, 10 -5 mol·L-1 insulin was significant,a large amount of cells died.10 -6 mol·L-1 insulin was effective and had high survival rate of HepG2 cells,which had sta-tistical significance compared with the normal group. When the incubation concentration of 2-NBDG was more than 100 mol·L-1 ,the fluorescence intensity of the cells was significantly different from the normal group.When the incubation time of 2-NBDG was more than 20 min,fluorescence intensity was significantly different from the normal group.When the incubation time of 2-NBDG was more than 100 min,the fluores-cence quenching phenomenon was obvious in the cells. Berberine hydrochloride and metformin significantly in-creased the glucose consumption and glucose uptake in cell supernatant, which had statistical significance compared with the model group.Conclusions Using 10 -6 mol · L-1 insulin induced IR model in HepG2 cells,the optimum incubation concentration and incu-bation time of 2-NBDG is 200 μmol·L-1 and 80 min, respectively.Berberine hydrochloride and metformin have obvious effect on improving IR in HepG2 cells.

Objective To investigate the effects and mechanisms of up-regulation of heme oxygenase-1(HO-1)on uric acid-induced adipocyte dysfunction. Methods Human bone marrow-derived mesenchymal stem cells(MSCs)were cultured in vitro and second or third generation MSCs were selected and recultured in an adipogenic induction medium,with the addition of various amounts of fructose and uric acid to find the optimal concentrations.Then,the HO-1 inducer cobalt protoporphyrin (CoPP)and the HO-1 inhibitor tin porphyrin(SnMP)were added successively.There were five independent samples in each group.Adipogenesis in MSC-derived adipocytes and droplets were measured by spectrophotometry,expression levels of xanthine oxidase(XO)and NADPH were measured by Western blott,superoxide levels were measured by Luminous spectrometer,and levels of adipogenic markers and HO-1 were measured by reverse transcription-polymerase chain reaction(RT-PCR). Results Fructose at 500 μmol/L and uric acid at 50 mg/L were the optimal concentrations for stimulating adipogenesis in MSC-derived adipocytes(P< 0.05).Compared with the controls,fructose significantly increased the levels of XO expression and uric acid(P< 0.05),and concurrent treatment with fructose and CoPP effectively reversed both XO and uric acid levels to those of the controls(all P<0.05).However,SnMP negated the beneficial effects of CoPP(P< 0.05).Additionally,uric acid decreased the number of small droplets and increased expression levels of adipogenic markers and NADPH(all P<0.05),but proadipogenic mediator levels were reduced with the addition of CoPP(all P<0.05).Furthermore,uric acid reduced expression levels of Wnt 10b,but had no significant effect on HO-1 expression,andthese effects were reversed upon the addition of CoPP(all P<0.05). Conclusions Upregulation of HO-1 can reduce the levels of XO and uric acid,adipogenesis in MSC-derived adipocytes,and also the expression of adipogenic markers and NADPH.Therefore,it plays a role in antioxidant stress.HO-1 agonists may be targets for treating organ damage and controlling weight in elderly obese patients with metabolic syndrome.

OBJECTIVETo compare the detection sensitivity of epidermal growth factor receptor (EGFR) mutations between allele specific oligonucleotide PCR (ASO-PCR) and bi-loop probe and specific primer quantitative PCR (BPSP-qPCR).

METHODSA total of 96 non-small cell lung cancer specimens were selected from West China Hospital from September 2009 to December 2010. ASO-PCR was developed to detect the presence of classical EGFR mutations. A total 39 available specimens were also tested by BPSP-qPCR.

RESULTSEGFR mutation detection rate was 30.2% (26/96) by ASO-PCR. The mutation rate was higher in female than in male patients [45.5% (20/44) vs. 17.3% (9/52), P = 0.003], non-smokers than smokers [44.1% (26/59) vs. 8.1% (3/37), P < 0.001] and adenocarcinomas than other subtypes of lung cancer [37.0% (27/73) vs. 8.7% (2/23), P = 0.01]. Among mutation negative cases by ASO-PCR, BPSP-qPCR increased the rate of detection of 19-del and L858R mutation by 10.3% (4/39) in adenocarcinomas and non-smoking subset. Overall, the mutation detection rate of BPSP-qPCR was higher than that of ASO-PCR [66.7% (26/39) vs. 41.0% (16/39), P = 0.02].

CONCLUSIONBPSP-qPCR has a better detection sensitivity than that of ASO-PCR.

Adenocarcinoma ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; DNA Mutational Analysis ; Female ; Genes, erbB-1 ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Mutation ; Polymerase Chain Reaction ; methods ; Receptor, Epidermal Growth Factor ; genetics ; Sensitivity and Specificity ; Sex Factors ; Smoking

OBJECTIVETo evaluate the quality of the radial artery for coronary artery bypass grafting (CABG) from patients with diabetes by observing the morphology of the radial artery and detecting the expression of vascular endothelial growth factor (VEGF) which may attribute to the long-term patency rate of the coronary artery bypass grafting.

METHODSSamples from 20 cases of diabetic and non-diabetic patients were prospective collected from June 2009 to December 2010. HE staining technique was used to test the morphology of radial artery through the observation of 20 cases of diabetic and 20 cases of non-diabetic patients who undergone CABG. The intimal thicken of the radial artery in the two groups of patients was compared. Western blot and immunofluorescence were then used to test the expression and location of VEGF in the two groups of patients.

RESULTSThe radial artery endothelial thickening index and intima/media ratio were significantly higher in the diabetic patients when compared with non-diabetic patients (0.90 ± 0.28 vs. 0.29 ± 0.25, t = 7.27, P < 0.01; 0.90 ± 0.21 vs. 0.37 ± 0.18, t = 8.57, P < 0.01). The expression of VEGF in diabetic patients was significantly higher than non-diabetic patients as revealed by Western blot (1.20 ± 0.21 vs. 0.67 ± 0.15, t = 6.49, P < 0.01). Immunofluorescence showed that VEGF distributed in the cytoplasm of the endothelial cells of diabetic patients radial artery.

CONCLUSIONSDiabetic patient's radial artery intimal thickness is significantly higher than non-diabetic patient's. VEGF may be an important inflammatory cytokine which is leading the radial artery intima thickening in the diabetic patients. The choice of the radial artery grafts in diabetic patients for CABG should be careful.

Aged ; Coronary Artery Bypass ; Coronary Artery Disease ; metabolism ; surgery ; Diabetes Mellitus ; pathology ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Radial Artery ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo observe the effects of bushen tongmai recipe (BSTMR) on mRNA and protein expressions of phosphatidylinositol-3-kinase (PI-3K) p85alpha in hepatic, adipose, muscular and ovarian tissues in PCOS rats with insulin resistance (IR).

METHODTwenty-three-day-old female SD rats were injected subcutaneously with sodium prasterone sulfate (90 mg x kg(-1) x d(-1)) for 20 days, and fed with high-fat forage for 80 days to induce PCOS rats with IR Then the rats were randomly divided into the model group and the treated group. Meanwhile, a group of fifteen rats of the same age was considered as the normal control group. The treated group were administered with BSTMR. The ovulation condition was examined by hematoxylin-eosin (HE) staining. Fasting blood glucose (FBG) was determined using glucose oxidase method. Serum fasting insulin (Fins) was determined by radioimmunoassay (RIA). The mRNA level of PI-3K p85alpha was measured by reverse transcription polymerase chain reaction(RT-PCR). Immunohistochemistry staining was wtilised to detect protein expression of PI-3K p85alpha in ovary.

RESULTCompared with the model group, the mean number of corpus luteum and the rate of ovulation in the treated group increased significantly (P <0. 01). The level of Fins in the treated group was much lower than that in the model group (P < 0.01). Both mRNA and protein expressions of PI-3K p85alpha in target tissues were up-regulated significantly in the treated group (P < 0.01 or P < 0.05).

CONCLUSIONBSTMR could improve IR and ovulation dysfunction in PCOS rats accompanying with IR and its molecular mechanisms might be closely related with the elevation of mRNA and protein levels of PI-3K p85alpha in target tissues of the model rats.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Gene Expression ; drug effects ; Humans ; Insulin Resistance ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Polycystic Ovary Syndrome ; drug therapy ; enzymology ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effects of Bushen Tongmai Recipe (, BSTMR) on mRNA and protein expressions of protein kinase B alpha (PKB alpha) in hepatic, adipose, muscular, and ovarian tissues of polycystic ovary (PCO) rats with insulin resistance (IR) and to explore the possible molecular mechanism of BSTMR in treating IR and ovulation dysfunction.

METHODSFemale 22-day-old SD rats were injected subcutaneously with sodium prasterone sulfate (9 mg.100g(-1).d(-1)) for 20 days and fed with high-fat diet for 80 days to induce PCO rats with IR. Then, the PCO rats were randomly divided into the model group (n=23) and the treated group (n=21). The treated group was administered with BSTMR for 2 weeks. Meanwhile, a group with 15 rats of the same age was used as the control group. The histological changes in the ovaries were examined. Fasting blood glucose (FBG) was determined by the glucose oxidase method. Serum fasting insulin (Fins) was determined by radioimmunoassay (RIA). The mRNA level of PKBalpha was measured by reverse transcription polymerase chain reaction (RT-PCR). Immunohistochemistry staining and Western blot analysis were employed to detect the protein expression in target tissues.

RESULTSCompared with the control group, the ovaries in the model group showed multiple follicular cysts, levels of FBG and Fins in the model group increased markedly (P<0.05 or P<0.01, respectively), and the insulin sensitive index (ISI) decreased obviously (P<0.01). The mRNA and protein expressions of PKBalpha in target tissues in the model group were dramatically lower than those in the control group (P<0.05 or P<0.01). Compared with the model group, the stratum granulosum of the ovarian follicle in the treated group increased markedly, the level of Fins in the treated group decreased obviously (P<0.01), ISI in the treated group improved markedly (P<0.01), and the mRNA and protein expressions of PKBalpha in target tissues of the treated rats were elevated significantly (P<0.05 or P<0.01).

CONCLUSIONBSTMR could improve IR and ovulation dysfunction in PCO rats with IR, and its molecular mechanisms might be closely related with the elevation of mRNA and protein expressions of PKBalpha in target tissues of PCO rats with IR.

Animals ; Blood Glucose ; drug effects ; metabolism ; Blotting, Western ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fasting ; blood ; Female ; Gene Expression Regulation, Enzymologic ; drug effects ; Insulin ; blood ; Insulin Resistance ; physiology ; Organ Specificity ; drug effects ; Ovary ; drug effects ; enzymology ; pathology ; Polycystic Ovary Syndrome ; blood ; drug therapy ; enzymology ; physiopathology ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To study the association between serum TB level and target organ damage in elderly metabolic syndrome (MS) patients.Methods Two hundred and forty-five elderly MS patients admitted to our hospital were included in this study.Their general condition was recorded,their serum TB,blood glucose,blood lipids,blood urea nitrogen and creatinine (Cr) levels were measured,and their left ventricular mass (LVM) and left ventricular mass index (LVMI) were detected by parallel echocardiography.Results Correlation analysis showed that the serum TB level was positively related with that of Cr (r=0.168,P=0.009) but not related with LVM,LVMI,serum blood urea nitrogen level,prevalence of chronic renal insufficiency and chronic kidney disease (P＞0.05).Regression analysis showed that serum TB level was an independent risk factor for urea in MS patients (OR=-0.27,95％CI:-0.48-0.06,P=0.01;OR=1.27,95％CI:0.33-2.22,P=0.01).Quantile analysis of serum TB level showed that the serum Cr level was significantly higher in Q76-100 group,Q51-75 group and Q26-50 group than in Q1-25 group (P=0.031).Conclusion Serum TB level is positively related with endogenous Cr clearance in elderly MS patients,suggesting that mildly elevated serum TB level may be a protective factor for impaired renal function in elderly MS patients.

Objective To evaluate the safety and efficacy of therapeutic ERCP for patients above 90 years of age.Methods The data of 37 patients of above 90 years who underwent 42 ERCP procedures from January 2001 to December 2009 were studied retrospectively and compared with those of 152 matched patients ( 168 procedures) below 65 years old at a 1∶4 ratio for success rate and complications.Results The rate of complete success,partial success,and failure in observation group was 73.81％ (31/42),19.05％(8/42) and 2.38％ (1/42),respectively,which were similar (P ＞0.05) with those in control group,with complete success rate at 85.12％ ( 143/168),partial success rate at 12.50％ (21/168) and failure rate at 2.38％ (4/168).The rate of terminated operation in observation group (4.76％,2/42) was significantly higher than that of the control group (0.00％,0,P =0.039).The overall rate of complication in observation group was 7.14％ ( 3/42 ),slightly higher than that of the control group ( 6.55％,11/168,P ＞0.05 ).There was no significant difference between the two groups regarding the rates and severity of such complications as pancreatitis,hemorrhage and infection ( P ＞ 0.05 ).No perforation or death was observed.Conclusion Therapeutic ERCP for patients of 90 years or older is safe and effective.Adverse events related to chronic concomitant diseases need early detection and proper management.

OBJECTIVEDuring 2003-2005, an outbreak of meningitis due to Neisseria meningitidis serogroup C occurred in China. With the aim to find strain clues result in the final epidemics, the ancestral strain 053442, a clinical isolate, and a carrier strain 053426 with different gene type were analyzed.

METHODSClinical strain 053442 and carrier strain 053426 were cultured on GC agar plates under the same condition. Two-dimensional electrophoresis was performed using the pH 3-10 nonlinear IPG strips of 24 cm length, and all the protein spots were identified by matrix-assisted laser desorption/ionization time of flight spectrometry.

RESULTS502 and 380 protein spots were identified in 053426 and 053442 respectively, relating to 266 and 202 different genes covering a wide range of cellular functions. The express volume and number of proteins involved in energy metabolism, protein synthesis and amino acid biosynthesis in 053426 were higher than in 053442. Virulence factor Opa, Opc and a series of proteins involved in pilus assembly and retraction were identified in 053442, which appear to be of primary importance in colonization and invasion of human cells. Compared to 053442, virulence protein species were less in 053426, with lower express volumes too. No Opa and Opc were detected in 053426.

CONCLUSIONSThe different protein expression profiles of the clinical strain 053442 and carrier strain 053426 in the present study provide some clues of the different pathogenicity of the two strains, which may account for result in the final epidemics.

Bacterial Proteins ; analysis ; Bacterial Typing Techniques ; China ; epidemiology ; Disease Outbreaks ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Meningitis, Meningococcal ; cerebrospinal fluid ; epidemiology ; microbiology ; Neisseria meningitidis, Serogroup C ; classification ; isolation & purification ; Proteome ; analysis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization