Objective To investigate the effect of Shenfu injectio on insulin resistance in rabbits undergoing cardiopulmonary bypass (CPB) . Methods Thirty New Zealand white rabbits of both sexes weighing 2.2-2.5 kg were randomly divided into 3 groups (n = 10 each): group I sham operation (group S); group Ⅰ CPB and group Ⅰ CPB + Shenfu injectio (group SFI). In group IE (group SFI) Shenfii injectio 10 ml/kg was injected intraperitoneally (IP) once a day for 2 days and at 30 min before anesthesia. In group Ⅱ and Ⅲ the duration between aortic cross-clamping and unclamping was 60 min. MAP was maintained at 50-60 mm Hg during CPB. The plasma glucose and insulin concentrations were measured immediately after induction of anesthesia (T_1), immediately after aortic cross-clamping (T_2) , and at 5, 35 and 75 min after aortic unclamping (T_3, T_4 , T_5) and insulin resistance index (HOMA-IR) was calculated. The expression of insulin receptor substrate-1 (IRS-1), phosphatidyl-inositol 3 kinase (PI3Kp85) and glucose transporters (CLUT4) in skeletal muscle was detected at 150 min of CPB by immuno-histochemistry. Results Compared with the baseline values at T_1 , plasma glucose and insulin concentrations and HOMA-IR were significantly increased at T_(2-5) in all 3 groups ( P < 0.05) . CPB significantly increased the plasma glucose and insulin concentrations and HOMA-IR at T_(2-5) in group Ⅱ and Ⅲ as compared with group S ( P < 0.05). The expression of PI3Kp85 and GLUT4 in muscle was significantly decreased in group Ⅱ and Ⅲ as compared with group S ( P < 0.05). Shenfu injectio significantly decreased plasma glucose concentration and HOMA-IR and increased plasma insulin concentration in group fD as compared with group Ⅱ . Shenfu injectio significantly attenuated the CPB-induced changes in IRS-1, PI3Kp85 and GLUT4 expression in muscle. Conclusion Shenfu injectio can ameliorate CPB-induced insulin resistance and the mechanism may be related to the up-regulation of the expression of insulin signal transduction molecules in skeletal muscle.

Objective To study the correlation between MMP-9 rs3918242 C ＞ T polymorphism and cancer risk.Methods PubMed database was utilized to search the relative artiles concerning association between MMP-9 rs3918242 C ＞T polymorphism and cancer risk (last search updated on April 2011).All the statistical analysis was performed by STATA software.Results 4,124 cancer patients and 4,728 control subjects were included in this meta-analysis.No association was observed in the overall analysis (allelic contrast P=0.378,OR =0.94,95％ CI:0.83-1.07).However,in the stratified analysis by cancer type,MMP-9-1562 T allele was found to decrease the lung cancer(P =0.026,OR =0.70,95％ CI:0.5 1-0.96) and colorectal cancer(P=0.016,OR =0.80,95％ CI:0.66-0.96).Conclusion MMP-9 rs3918242 C/T polymorphism is correlated with colorectal cancer and lung cancer.

OBJECTIVETo obtain the information on the glucuronidation of Ginkgo flavonoid and the interaction profile of Ginkgo flavones with other drugs in vitro.

METHODSGinkgo flavonoids (quercetin, isorhamnetin and keampferol) and other drugs were co-incubated with rat hepatic microsome at 25 degree; the residual concentrations of flavonoids were determined by HPLC. The enzymatic parameters of quercetin, isorhamnetin and keampferol metabolism were assessed. The interactions between flavonoids and these drugs on glucuronidation were observed.

RESULTThe K(m) values were ( 24+/-0.05), (148+/-0.09) and (110+/-0.03) micromol/L and the V(max) values were (60+/-0.21), (48+/-0.02) and (34+/-0.02) micromol x g(-1) x min(-1) for quercetin, isorhamnetin and kaempferol, respectively. The IC(50) of nifedipine propafenone ipriflavone and diphenytriazol on flavonoids metabolism were 54 - 70, 69 - 122, 85 - 98 and 210 - 362 micromol, respectively. The inhibition constants (Ki) of diphenytriazol propafenone and ipriflavone on quercetin, isorhamnetin and keampferol metabolism were (57.6, 50.5, 33.1) (33.6, 59.5, 45.2) and(13.7,24.0,15.7) microg/ml respectively. The ratio [I]/[Ki] of the plasma concentration and inhibition constant for propafenone was 0.002 - 0.003.

CONCLUSIONThe metabolic level of quercetin is the strongest among three Ginkgo flavonoids. Nifedipine propafenone and ipriflavone inhibit the metabolism of quercetin, isorhamnetin and keampferol at different levels. Because of the interaction between Ginkgo flavonoids with nifedipine, caution must be taken when two drugs are used together clinically.

Animals ; Drug Interactions ; Flavonoids ; metabolism ; Ginkgo biloba ; Glucuronides ; metabolism ; Microsomes, Liver ; metabolism ; Rats

BACKGROUND: Resistin has been shown to play an important role in the pathogenesis and progression of osteoarthritis. OBJECTIVE: To investigate the correlation between resistin level in serum and synovial fluid and the severity of knee osteoarthritis. METHODS: Seventy-four patients with knee osteoarthritis and 79 healthy individuals were recruited. The Noyes score method was used to assess articular cartilage damage arthroscopically. The severity of knee osteoarthritis was evaluated according to the Western Ontario McMaster University Osteoarthritis (WOMAC) score. The radiographic severity of knee osteoarthritis was assessed by the Kellgren-Lawrence (K-L) grading system. The levels of resistin and cross linked C-telopeptide of type Ⅱ collagen (CTX-Ⅱ) in serum and synovial fluid were determined by ELISA. RESULTS AND CONCLUSION: The resistin level in the synovial fluid was positively correlated with the radiographic severity of knee osteoarthritis, WOMAC pain, WOMAC function, and WOMAC total scores, but showed no correlation with the WOMAC stiffness score. There were no significant correlations between the serum level of resistin and WOMAC total scores, subscale or K-L Grading system in the the osteoarthritis patients. Multiple regression analysis showed that the correlation between the synovial fluid level of resistin and WOMAC scores was of significance after removal of other confounding factors, and the resistin level in the synovial fluid was positively correlated with Noyes scores and CTX-Ⅱ level. These results suggest that the resistin level in the synovial fluid can serve as a potential biomarker reflecting the severity of knee osteoarthritis and cartilage degenerative degree in patients with osteoarthritis.

Objective To investigate surgical treatment and effect of secondary endocardial fibroelastosis,based on respective analysis of clinical data and follow-up data of patients with secondary endocardial fibroelastosis (SEF) between 2010 and 2012.Methods A retrospective analysis was performed including 10 patients with secondary endocardial fibroelastosis from January 2010 to December 2012 in Wuhan Union Hospital.All patients were diagnosed by Untrasonic Cardiogram and/or CT angiography of heart and great vessel,and had cardiac insufficiency in different degree [EF 0.37 ± 0.08 (0.26 ～ 0.48)].All patients except 2 patients with anomalous origin of the coronary artery received treatment of digitaloid drugs before operation,which promoted preoperative cardiac function.5 patients with SEF complicated with Congenital Coarctation of the Aorta (CoA),2 patients underwent correction of CoA,2 patients underwent correction of CoA and partial resection of endocardium,1 patient underwent correction of CoA,partial resection of endocardium and mitral vavuloplasty.2 patients with SEF complicated with anomalous origin of the left coronary artery from the pulmonary artery,who were underwent correction of anomalous origin of coronary artery.2 patients with SEF complicated with aortic stenosis,who were underwent aortic commissurotomy and partial resection of endocardium.1 patient with SEF complicated with mitral stenosis and insufficiency,who underwent mitral valve replacement.The intraopertive gross appearance of endocardium was opaque greyish-white not transparent pink.The postoperative pathological examination showed obviously positive dyeing of elastic fibers.In 3,6,12 and 24 months after operation,Untrasonic Cardiogram evaluated cardiac function and endocardium.Results one 6 months patients with origin of left coronary from pulmonary artery died of severe post-operative low cardiac output syndrome,while another 1 months patients with origin of left coronary from pulmonary artery obtained post-operative good recovery,and the Untrasonic Cardiogram show disappearance of endocardial fibroelastosis.The post-operative mean time of using respirator(4.0 ± 1.5) days (2-7 days).Compared with the preoperative data,the cardiac function index (EF) was not significantly better at 2 weeks and 3-6 months[0.38 ± 0.07 (0.28 ～ 0.48),P ＞ 0.05 ; 0.39 ± 0.08 (0.30 ～ 0.50),P ＞ 0.05],and the non-resected fibroelatic endocardium still existed and were not attenuated.But the cardiac function index (EF) significantly increased [0.44 ± 0.08 (0.38 ～ 0.55),P ＜ 0.05] than the pre-operative EF,and the 3 of 5 cases the fibroelatic endocardium were attenuated or disappeared,while 2 of 5 cases the fibroelatic endocardium still existed.Conclusion SEF is the important causes of the infant intractable heart failure,which has the characteristic of high mortality and limited therapy.For SEF patients with anomalous origin of the coronary artery,the SEF is completely reversed by early diagnosis and early correction of the malformation.For SEF patients with CoA or aortic stenosis,the surgical treatment could promote recovery of cardiac function,but whether the SEF were reversed is still subject to further follow-up.The heart transplantation is the best therapy for SEF with severse heart failure.

Objective To analyze the characteristic colligation force vector curves in hemiplegic patients. Methods Eleven hemiplegic patients were recruited, and 11 healthy elderly served as control. The characteristic changes of the patients with regard to colligation force vector demonstrated on a feedback apparatus of the "X.O.S"system during the dynamic training exercise program were analyzed and compared with that of the control group. Results The value of the colligation force vector passing most of the regions were significantly decreased ( P

To clone the 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase (TwMCT) full length cDNA from Tripterygium wilfordii, the specific primers were designed according to the transcriptome data and the LCPCR were carried out. After a series of bioinformatics analysis on the TwMCT, the MeJA induced expression content were investigated by real-time fluorescence quantification polymerase chain reaction (RT-qPCR). The result showed that the full of TwMCTcDNA was 1 318 bp nucleotides encoding 311 amino acids. The molecular weight of the deduced TwMCT protein was about 34.14 kDa and the theoretical isoelectric point was 8.65. Result of the RT-qPCR analysis indicated that the content of TwMCT mRNA expression in T. wilfordii suspension cell was rising after treating with MeJA and reached the maximum in 24 h. Cloning and analyzing TwMCT gene from T. wilfordii provided gene element for studying the function and expression regulation of secondary metabolites.
Amino Acid Sequence ; Cloning, Molecular ; Erythritol ; analogs & derivatives ; metabolism ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Nucleotidyltransferases ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary ; Sequence Alignment ; Sugar Phosphates ; metabolism ; Tripterygium ; chemistry ; enzymology ; genetics

AIMTo develop a method for assaying Ginkgo flavones in rat hepatical microsome.

METHODSQuercetin, isorhamnetin and keampferol were added to microsome incubate and incubated for a given time then extracted with ether-acetone. After evaporated, the residue was reconstituted with 100 microL of phosphate buffer solution (pH 2.0)-tetrahydrofuran-methanol-isopropanol (60:15:10:20). An aliquot of 20 microL was injected into the HPLC system. According to the result of estimate by means of HPLC, the results of metabolism of Ginkgo flavones in different conditions was compared.

RESULTSThe assay was linear over the rang of 0.2-8 mg.L-1 for Ginkgo flavones. The limit of quantification was 0.1 mg.L-1 (n = 3). The recoveries of three components of Ginkgo flavones were 99.9%-113.8% for quercetin (RSD < 0.8%), 100.8%-117.3% for isorhamnetin (RSD < 1.9%) and 100.7%-116.5% for keampferol (RSD < 1.03%, n = 5).

CONCLUSIONThe method is simple, fast and accurate. It can be used for investigation of the metabolism of Ginkgo flavones.

Animals ; Chromatography, High Pressure Liquid ; Flavonols ; isolation & purification ; metabolism ; Ginkgo biloba ; chemistry ; In Vitro Techniques ; Kaempferols ; isolation & purification ; metabolism ; Microsomes, Liver ; metabolism ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; isolation & purification ; metabolism ; Rats

OBJECTIVETo obtain recombinant human CYP2E1 and to determine its activity by using the specific probe substrate.

METHODSCYP2E1 cDNA was obtained by RT-PCR using human liver RNA as template. The cloned CYP2E1 cDNA was ligated with pFastBac vector to generate recombinant pFastBac-CYP2E1, which was then transformed into E. coli DH 10 Bac. Recombinant Bacmid-CYP2E1 was generated by transposition. Then Spodoptera frugiperda (Sf9) insect cells was infected with Bacmid-CYP2E1 to generate recombinant baculoviruses carrying human CYP2E1 cDNA. Finally, Sf9 insect cells were triinfected with recombinant baculoviruses carrying human CYP2E1, CYPOR and CYPb5. The activity of the recombinant enzymes was determined using chlorzoxazone as the substrate.

RESULTThe Kmand Vmaxof recombinant CYP2E1 to chlorzoxazone was (72.4 +/-8.7) micromol. L(-1) and (2.41 +/-0.10) micromol.min(-1)?g(-1)protein, respectively.

CONCLUSIONActive recombinant CYP2E1 has been obtained by bac-to-bac expression system and its activity is similar to previous reports.

Animals ; Baculoviridae ; genetics ; metabolism ; Cytochrome P-450 CYP2E1 ; biosynthesis ; genetics ; Escherichia coli ; genetics ; Genetic Vectors ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Spodoptera ; genetics ; metabolism ; Transfection

OBJECTIVE: To evaluate enantiomeric separation methods for beta-blocking agents and analogs. METHODS: Enantiomeric separation of racemates of 11 beta-blocking agents and their analogs was performed using chiral stationary phases and 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl isothiocyanate (GITC). RESULTS: These beta -blocker racemates were separated into enantiomers in one or several chormatographic states such as propranolol, bisoprolol, metoprolol, celiprolol, carvedilol, sotalol, propafenone, ephedrine, and zomitriptan. Temperature had a significant effect on the resolution of the drugs when using chiralcel OD. Lower temperatures were associated with higher resolutions. CONCLUSION: When separating beta-blocking agents and their analogs, Chiralcel OD, Chiralpak AD, Chiral stationary phases and GITC chiral derivative reagents have complementary functions.