The metabolic characteristics of ligustrazin (TMPz) in liver microsomes were investigated in the present study. The reaction phenotyping of TMPz metabolism was also identified by in vitro assessment using recombinant human cytochrome P450 enzymes (CYP) and UDP glucuronosyltransferases (UGT). TMPz was incubated at 37 degrees C with human (HLM) and rat liver microsomes (RLM) in the presence of different co-factors. The metabolic stability and enzyme kinetics of TMPz were studied by determining its remaining concentrations with a LC-MS/MS method. TMPz was only metabolically eliminated in the microsomes with NADPH or NADPH+UDPGA. In the HLM and RLM with NADPH+UDPGA, t1/2, K(m) and V(max) of TMPz were 94.24 +/- 4.53 and 105.07 +/- 9.44 min, 22.74 +/- 1.89 and 33.09 +/- 2.74 micromol x L(-1), 253.50 +/- 10.06 and 190.40 +/- 8.35 nmol x min(-1) x mg(-1) (protein), respectively. TMPz showed a slightly higher metabolic rate in HLM than that in RLM. Its primary oxidative metabolites, 2-hydroxymethyl-3, 5, 6-trimethylpyrazine (HTMP), could undergo glucuronide conjugation. The CYP reaction phenotyping of TMPz metabolism was identified using a panel of recombinant CYP isoforms (rCYP) and specific CYP inhibitors in HLM. CYP1A2, 2C9 and 3A4 were found to be the major CYP isoforms involved in TMPz metabolism. Their individual contributions were assessed b) using the method of the total normalized rate to be 19.32%, 27.79% and 52.90%, respectively. It was observed that these CYP isoforms mediated the formation of HTMP in rCYP incubation. The UGT reaction phenotyping of HTMP glucuronidation was also investigated preliminarily by using a panel of 6 UGT isoforms (rUGT). UGT1A1, 1A4 and 1A6 were the predominant isoforms mediated the HTMP glucuronidation. The results above indicate that the metabolism of TMPz involves multiple enzymes mediated phase I and phase II reactions.

AIMTo investigate the effects of the recombina nt human endostatin on adjuvant arthritis (AA) in rats and its mechanisms. METHODSThe model of rat AA was induced by injection of intradermal CFA. ConA and LPS induce d splencytes proliferation was examined by MTT asssy and the activities of inter leukin-1 (IL-1) and IL-2 were measured by the method of thymocytes prolifera tion. Synoviocytes were dispersed with incubation of collagenase and trypsin, an d IL-1, TNF production of synoviocytes was estimated with radio-immunity assay . RESULTSThe secondary inflammation of AA rats appeared on the 1 0th day after injection of CFA. The therapeutic administration of endostatin (0 1, 0 5, 2 5 mg?kg -1 ?d -1 , sc, ?7 d) was given at that time(d 10). It was found that endostatin significantly inhibited the secondary paw swel ling. The increased ConA-induced splenic lymphocyte proliferation reaction and the activated IL-1 and IL-2 activity of AA rats was reversed by the treatment with endostatin. Meanwhile,IL-1 produced by PM? was increased. Endostatin inh ibited IL-1 production from PM? and IL-1 and reduced TNF level of Synoviocyte s in AA rats. CONCLUSIONThe recombinant human endostatin has the rapeutical effect on AA rats, its mechanisms is related to its immunoregulative function.

24 hours) and versus control group(

The metabolic characteristics of ligustrazin (TMPz) in liver microsomes were investigated in the present study. The reaction phenotyping of TMPz metabolism was also identified by in vitro assessment using recombinant human cytochrome P450 enzymes (CYP) and UDP glucuronosyltransferases (UGT). TMPz was incubated at 37 degrees C with human (HLM) and rat liver microsomes (RLM) in the presence of different co-factors. The metabolic stability and enzyme kinetics of TMPz were studied by determining its remaining concentrations with a LC-MS/MS method. TMPz was only metabolically eliminated in the microsomes with NADPH or NADPH+UDPGA. In the HLM and RLM with NADPH+UDPGA, t1/2, K(m) and V(max) of TMPz were 94.24 +/- 4.53 and 105.07 +/- 9.44 min, 22.74 +/- 1.89 and 33.09 +/- 2.74 micromol x L(-1), 253.50 +/- 10.06 and 190.40 +/- 8.35 nmol x min(-1) x mg(-1) (protein), respectively. TMPz showed a slightly higher metabolic rate in HLM than that in RLM. Its primary oxidative metabolites, 2-hydroxymethyl-3, 5, 6-trimethylpyrazine (HTMP), could undergo glucuronide conjugation. The CYP reaction phenotyping of TMPz metabolism was identified using a panel of recombinant CYP isoforms (rCYP) and specific CYP inhibitors in HLM. CYP1A2, 2C9 and 3A4 were found to be the major CYP isoforms involved in TMPz metabolism. Their individual contributions were assessed b) using the method of the total normalized rate to be 19.32%, 27.79% and 52.90%, respectively. It was observed that these CYP isoforms mediated the formation of HTMP in rCYP incubation. The UGT reaction phenotyping of HTMP glucuronidation was also investigated preliminarily by using a panel of 6 UGT isoforms (rUGT). UGT1A1, 1A4 and 1A6 were the predominant isoforms mediated the HTMP glucuronidation. The results above indicate that the metabolism of TMPz involves multiple enzymes mediated phase I and phase II reactions.
Animals ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP2C9 ; metabolism ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Drug Interactions ; Glucuronosyltransferase ; metabolism ; Humans ; Ligusticum ; chemistry ; Microsomes, Liver ; enzymology ; NADP ; metabolism ; pharmacology ; Pyrazines ; metabolism ; pharmacokinetics ; Rats ; Uridine Diphosphate Glucuronic Acid ; metabolism ; pharmacology

Objective To investigate the early clinical outcome of peripheral cutting balloon(PCB) in the management of peripheral vessel stenosis.Methods Thirteen patients with peripheral limb vessel stenosis, in which 4 stenoses in hemodialysis access and 9 at lower limb arteries,underwent angioplasty by PCB.For multiple stenosis in the same vessel,the distal one should be expanded firstly.The balloon pressure was controlled in the range of 8 atm to 10 atm(1 atm=10.108 kPa).All the patient were given continuous anticoagulant therapy after the procedure.Results All the procedure were carried out successfully on the 13 patients,and no serious complications occured.The symptoms did not recur in all patients after the procedure.The 5 months' follow-up angiography proved that no restenosis occurred in one patient with previous stenosis at the hemodialysis access.Conclusion The angioplasty with PCB was a safe and reliable procedure in management of the peripheral limb vessel stenosis.The early outcome is satisfying.

AIM: To evaluate the efficiency and complications of single lacrimal duct intubation versus annular lacrimal duct intubation with 5-fluorouracil and tobramycin/dexamethasone eye ointment injection in lacrimal duct obstruction treatment.METHODS:A total of 74 patients (92 eyes) with lacrimal duct obstruction who received surgical treatment by Department of Ophthalmology in Qinhuangdao Haigang Hospital were consecutively recruited between August 2015 and September 2016.They were randomly allocated to Group A or Group B.After probing of lacrimal passage, Group A (46 eyes) were treated by using single lacrimal duct intubation.Group B (46 eyes) were treated by using annular lacrimal duct intubation combined with 5-fluorouracil and tobramycin/dexamethasone eye ointment injection.Between the two groups, Mann-Whitney Rank sum test was used to comparing the operative effect, and Chi-square test was used to comparing the occurrence of complications.RESULTS:The cure rate was 61% in Group A and 89% in Group B;the improvement rate was 22% in Group A and 4% in Group B.Group B had better effect and less complications (2 cases) than Group A (8 cases),the differences between the two groups were significantly(P<0.05).CONCLUSION: Compared with single lacrimal duct intubation, annular lacrimal duct intubation combined with 5-fluorouracil and tobramycin/dexamethasone eye ointment injection has better operative effect and less complication in lacrimal duct obstruction treatment.

AIM: To evaluate the operative effect and time effectiveness of the conventional surgery versus retrograde gas injection under nasal endoscope combined 5-Fluorouracil for lacrimal canalicular rupture.
●METHODS:A total of 67 patients (67 eyes) with lower lacrimal canalicular rupture who received surgical treatment by Department of Ophthalmology in Qinhuangdao Haigang Hospital were consecutively recruited between Jan. 2009 and Dec. 2015. They were randomly divided into Group A or Group B. Group A (33 patients, 33 eyes) were treated by conventional surgery, and Group B ( 34 patients, 34 eyes ) were treated by retrograde gas injection under nasal endoscope combined 5-Fluorouracil. Time for finding out the cute end of the lower lacrimal canaliculus and postoperative effect were recorded. Comparisons between the two groups were done with lndependent sample t-test and Mann-Whitney Rank sum test.
●RESULTS: Time for finding out the cute end of the lower lacrimal canaliculus of Group A was (44. 42±10. 66) min, and the time of Group B was ( 30. 06 ± 6. 21 ) min. There was significant difference between the two groups (t=6. 72, P<0. 05). Lacrimal ducts flush was done at the 6mo after the survey, Group B had better effect than Group A, the difference between the two groups were significant (Z=2. 47,P<0. 05).
●CONCLUSION: Compared with conventional surgery, retrograde gas injection under nasal endoscope combined 5-Fluorouracil for lacrimal canalicular rupture can make the operation time shorter and has better effect.

AIMTo investigate the effects of interleukin-2 (IL-2) on the transmembrane potential and contractile force in ventricular papillary muscle of rat and the underlying mechanism.

METHODSThe transmembrane potentials and contractile force were recorded by intracellular glass microelectrode and tension transducer in the isolated rat papillary muscles.

RESULTS(1) IL-2 shortened the action potential duration (APD50 and APD80), while had no effects on resting potential, action potential amplitude and depolarization rate. (2) IL-2 depressed the contractile force of the muscle in dose-dependent manner. IL-2 at concentrations of 0.5, 2.5, 10, 50 and 200 u/ml decreased the developed tension to 94.8% (P < 0.05), 85.8%, 76.3%, 69.3% and 52.5% (P < 0.01), respectively. (3) Pretreatment with L-NAME (10(-4) mol/L) attenuated the negative inotropic effect of IL-2, in which effect of IL-2 at concentrations from 0.5 to 10 u/ml was completely abolished, and the effect of IL-2 at high dose (50 and 200 u/ml) was partly attenuated by L-NAME.

CONCLUSIONIL-2 had inhibitory effects on action potential duration and contractile force of papillary muscle, and its negative inotropic effect was mediated by nitric oxide.

Action Potentials ; drug effects ; Animals ; Heart Ventricles ; drug effects ; Interleukin-2 ; pharmacology ; Male ; Myocardial Contraction ; drug effects ; physiology ; Nitric Oxide ; metabolism ; Papillary Muscles ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley

With the establishment of HPLC and LC-MS methods to determine the related substances and the content of active pharmaceutical ingredient (API) in ipratropium bromide aerosol products, several packing material-related impurities were identified, including antioxygen BHT and antioxygen 2246. Results showed that these leachable additives from the packing materials may present at a relative high level in the drug solution, and the low content of API in the drug products is usually due to the adsorption of the packing material as well as the leaking of contents. The current available assay methods for the control of ipratropium bromide aerosol products are often lack of specificity and unable to assure the drug quality effectively. To meet the increasing attention on the regulations of drug packing materials, our research would be a pilot study, indicating that the inappropriate packing materials could cause the migration and adsorption of the active ingredients, and the importance to have compatibility studies between packing materials and drugs.
Aerosols ; Antioxidants ; analysis ; Bronchodilator Agents ; administration & dosage ; chemistry ; Butylated Hydroxytoluene ; analysis ; Chromatography, High Pressure Liquid ; Drug Incompatibility ; Drug Packaging ; Ipratropium ; administration & dosage ; chemistry ; Quality Control ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

In-vitro assay methods were established to evaluate transactivation and binding activity of compounds on peroxisome proliferator-activated receptor y (PPARγ). Firstly, plasmids were constructed for transactivation assay of PPARγ response element (PPRE) triggered reporter gene expression, and for cell-based binding activity assay of the chimeric receptor, which was fused with PPARγ ligand binding domain (LBD) and yeast transcriptional activator Gal4. Secondly, by using PPARy competitive binding assay based on time resolved-fluorescence resonance energy transfer (TR-FRET), affinities of compounds and drugs to PPARγ were evaluated. In application of these above methods, the PPARγ activating potency and characteristics of different compounds were evaluated, and a novel benzeneselfonamide derivative, ZLJ01, was found to have comparable binding activity and affinity with the well-known PPARy agonist, but lack of PPRE mediated transactivation activity. In preliminary study on in-vitro hypoglycemic activity, ZLJ1 was found to promote insulin-stimulated glucose uptake by liver cells. Therefore, we believe that combining transactivation and binding activity as well as affinity evaluation, the system could be used to screen non-agonist PPARγ ligand as anovel PPARγ modulator
Genes, Reporter ; Hepatocytes ; Hypoglycemic Agents ; chemistry ; Ligands ; PPAR gamma ; agonists ; chemistry ; Plasmids ; Response Elements ; Sulfonamides ; chemistry ; Transcriptional Activation