The inter-species differences of thienorphine metabolism were investigated in human, Beagle dog and rat liver microsomes, by comparing enzyme kinetics of the parent drug and the formation of its major metabolites. The incubation systems of thienorphine with liver microsomes of the three species were optimized in terms of thienorphine concentration, microsomal protein content and incubation time. The concentrations of thienorphine and its metabolites in incubates were measured by a LC-MS/MS method. The biotransformation of thienorphine by human liver microsomes was the lowest among the three species. The K(m), V(max), CL(int) and T1/2 of thienorphine obtained from human liver microsomes were (4.00 +/- 0.59) micromol x L(-1), (0.21 +/- 0.06) micromol x L(-1) x min(-1), (117 +/- 3.19) mL x min(-1) x kg(-1) and (223 +/- 6.10) min, respectively. The corresponding kinetic parameters for dog and rat liver microsomes were (3.57 +/- 0.69) and (3.28 +/- 0.50) micromol x L(-1), (0.18 +/- 0.04) and (0.14 +/- 0.04) micromol x L(-1) x min(-1), (213 +/- 1.06) and (527 +/- 7.79) mL x min(-1) x kg(-1), (244 +/- 1.21) and (70.7 +/- 1.05) min, respectively. A total of six phase I metabolites were observed in liver microsomes, including one N-dealkylated metabolite, three oxidative metabolites and two N-dealkylated oxidation metabolites. All these six metabolites were detected in the liver microsomes of the three species. However, the relative amounts of the metabolites generated were different in three species. The results indicated that the major phase I metabolic pathway of thienorphine was similar in the liver microsomes from all three species. However, the inter-species differences observed were relative amounts of the metabolites as well as the metabolic characteristics of thienorphine in liver microsomal incubates.

The inter-species differences of thienorphine metabolism were investigated in human, Beagle dog and rat liver microsomes, by comparing enzyme kinetics of the parent drug and the formation of its major metabolites. The incubation systems of thienorphine with liver microsomes of the three species were optimized in terms of thienorphine concentration, microsomal protein content and incubation time. The concentrations of thienorphine and its metabolites in incubates were measured by a LC-MS/MS method. The biotransformation of thienorphine by human liver microsomes was the lowest among the three species. The Km, Vmax, CLint and T1/2 of thienorphine obtained from human liver microsomes were (4.00 ? 0.59) ?mol?L-1, (0.21 ? 0.06) ?mol?L-1?min-1, (117 ? 3.19) mL?min-1?kg-1 and (223 ? 6.10) min, respectively. The corresponding kinetic parameters for dog and rat liver microsomes were (3.57 ? 0.69) and (3.28 ? 0.50) ?mol?L-1, (0.18 ? 0.04) and (0.14 ? 0.04) ?mol?L-1?min-1, (213 ? 1.06) and (527 ? 7.79) mL?min-1?kg-1, (244 ? 1.21) and (70.7 ? 1.05) min, respectively. A total of six phase I metabolites were observed in liver microsomes, including one N-dealkylated metabolite, three oxidative metabolites and two N-dealkylated oxidation metabolites. All these six metabolites were detected in the liver microsomes of the three species. However, the relative amounts of the metabolites generated were different in three species. The results indicated that the major phase I metabolic pathway of thienorphine was similar in the liver microsomes from all three species. However, the inter-species differencesobserved were relative amounts of the metabolites as well as the metabolic characteristics of thienorphine in liver microsomal incubates.

BACKGROUND: The association between the ingredients of perfusate and its protection on corneal endothelium is always the hot issue in ophthalmology and pharmacology.OBJECTIVE: To observe the influence of different perfusates on the structure and function of rabbit corneal endothelium during phacoemulsification.DESIGN: A randomized grouping designed and controlled animal trial.SETTINGS: Laboratory of experimental animal center of Jinzhou Medical College and the laboratory of experimental animal operation of an urban hospital.MATERIALS: The experiments were carried out in the laboratory of experimental animal center of Jinzhou Medical College and the laboratory of experimental animal operation of Jinzhou Yadong Ophthalmology Hospital from September 2004 to March 2005. Sixteen pure Japanese big-ear rabbits of 3.5 months old, clean degree, were randomly divided into four groups with 4 rabbits in each group: normal control group, saline group, shike group and balanced salt solution group. Shike was produced by Shenyang Qixing Pharmaceutical Co.,Ltd.; Balanced salt solution by Alcon Company (USA).METHODS: The rabbits were intraperitoneally anesthetized with 100 ml/L chloral hydrate (3 mL/kg), 4% oxybuprocaine hydrochloride eye drops were used for surface anesthesia of eyes, and both eyes were operated. Alcon phacoemulsification apparatus (USA) and routine microsurgical instruments were used. A 3.5-mm incision was made on sclerotic tunnel at 2 mm posterior to superior limbus of sclera, punctured into the anterior chamber, then 0.25 mL Viscoat (Alcon) was infused. Curvilinear capsulorhexis was performed with the diameter of about 5 mm. The phacoemulsification head was placed in the center to suck out the crystal nucleus and cortex, and the incision was closed after the operation. The morphology of the corneal endothelium was quantitatively determined using contact specular microscope preoperativley and 6 hours postoperatively, including the density and area of corneal endothelium,percent of hexagonal cells and the coefficient of variation. At 6 hours postoperatively, trypan blue-alizarin red active staining was performed, and the changes of slight structures of corneal endothelium were observed under light microscope.MATN OUTCOME MEASURES: ① Quantitative analysis of the forms morphology of corneal endothelium (density and area of corneal endothelium, percent of hexagonal cells and the coefficient of variation); ② Characters of forms and structures of corneal endothelium.RESULTS: All the 16 rabbits were involved in the analysis of results. ① There were no significant differences in the morphological indexes among the four groups preoperatively. ② At 6 hours postoperatively, density and areas of endothelial cells, percent of hexagonal cells and the coefficient of variation were significantly lower in the saline group,shike group and balanced salt solution group than in the normal control group (P ＜ 0.05), and no significant difference was observed between the shike group and balanced salt solution group (P ＞ 0.05). ③ The structural changes of corneal endothelium in the shike group and in balanced salt solution group were alleviated more significantly than those in the saline control group, no necrosis was observed.CONCLUSTON: In phacoemulsification, the damage of perfusate to corneal endothelium is a chemical one. Under the same surgerical conditions, domestic perfusate of shike is as effective as balanced salt solution in protecting endothelial cells.

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.

Rotundine (1 micromol L(-1)) was incubated with a panel of rCYP enzymes (1A2, 2C9, 2C19, 2D6 and 3A4) in vitro. The remained parent drug in incubates was quantitatively analyzed by an Agilent LC-MS. CYP2C19, 3A4 and 2D6 were identified to be the isoenzymes involved in the metabolism of rotundine. The individual contributions of CYP2C19, 3A4 and 2D6 to the rotundine metabolism were assessed using the method of total normalized rate to be 31.46%, 60.37% and 8.17%, respectively. The metabolites of rotundine in incubates were screened with ESI-MS at selected ion mode, and were further identified using MS2 spectra and precise molecular mass obtained from an Agilent LC/Q-TOF-MSMS, as well as MS(n) spectra of LC-iTrap-MS(n). The predominant metabolic pathway of rotundine in rCYP incubates was O-demethylation. A total 5 metabolites were identified including 4 isomerides of mono demethylated rotundine and one di-demethylated metabolite. The results also showed that CYP2C19, 2D6 and 3A4 mediated O-demethylation of methoxyl groups at different positions of rotundine. Furthermore, the ESI-MS cleavage patterns of rotundine and its metabolites were explored by using LC/Q-TOF-MSMS and LC/iTrap-MS(n) techniques.

Angiotensin II (ANGII) plays an important role in the pathogenesis of atherosclerosis by inducing proliferation of vascular smooth muscle cells (VSMCs). In our study, we observed the effects of valsartan on proliferation of cultured VSMCs treated with or without ANGII by cell counting and methyl thiazolyl tetrazolium (MTT) assay, and detected the expression of mitofusin 2 (Mfn2), a newly discovered cell proliferation inhibitor and a related cell proliferation signaling pathway protein by Western blotting. ANGII at a concentration of 10(-6) mol/L significantly stimulated VSMCs proliferation, down-regulated the expression of Mfn2 and up-regulated the expression of Raf and ERK1/2. Valsartan inhibited such effects of ANGII at concentrations of 10(-5) and 10(-6) mol/L, but not at 10(-7) mol/L. Valsartan had no significant effect on the proliferation of untreated VSMCs. These results suggest that valsartan inhibits ANGII-induced proliferation of VSMCs in vitro via Mfn2-Ras-Raf-ERK/MAPK signaling pathway.

Aim To investigate the inhibitory and in-duction effects of the active components of SIWU de-coction on cytochrome P450 enzymes ( CYP ) and as-sess the CYP based drug interaction. Methods Ac-tive components fructose, ferulic acid, paeoniflorin, li-gustrazine and their combinations were incubated sepa-rately with human liver microsomes ( HLM) and probe substrates. Metabolites of the CYP probe substrates were determined by LC-MS/MS to assess the inhibitory activities on human CYP1 A2 , 2 B6 , 2 C9 , 2 C19 , 2 D6 and 3 A4 . Sandwich-cultured rat hepatocytes model was used to evaluate the CYP1 A2 and CYP3 A1/2 induc-tion. Results The inhibitory rates on CYP1A2, 2B6, 2 C9 and 2 C9 by the test groups at 100 μmol · L-1 were all < 62%, while the activities of CYP3 A4 and 2D6 were not affected. The CYP1A2 activities in the test groups of peoniflorin and its combinations ( 50μmol·L-1 ) were significantly enhanced, with the in-creasing fold more than 40% of positive control group. No significant induction on rat CYP3 A1/2 was ob-served for four principles and their combinations. Con-clusions The active components of SIWU decoction do not show significant inhibitory effects on six CYP isoforms. Peoniflorin could induce the CYP1A2 activity in rat hepatocytes. The induction activity is enhanced by the concomitant use of peoniflorin with ferulic acid and/or ligustrazine.

Objective To evaluate the clinical value of 99Tcm-MIBI scintigraphy for diagnosis of residual thyroid tissue and metastasis in patients with DTC after their first 131I therapy.Methods From February 2010 to March 2014,192 DTC patients (38 males,154 females,average age (43.2±8.6) years) who received total or near-total thyroidectomy and were pathologically diagnosed as DTC (171 papillary and 21 follicular carcinomas) underwent 99Tcm-MIBI scan (average dosage:740-925 MBq) 6 months after their first 131 I therapy.131 I scans was performed 4 d after oral administration of 131I of therapeutic dose (average dosage:5 550-8 140 MBq).Pre-and post-therapeutic images and the serum Tg level (detected before the imagings) were compared and analyzed.Any abnormal uptake of agent found inside or outside the thyroid was regarded as positive result.Patient-based and lesion-based data analysis were performed by x2 test and two-sample t test.Results A total of 191 patients were finally included,of which 65 positive cases were found.The sensitivity of 99Tcm-MIBI imaging was significantly lower than that of 131I imaging(56.9％ (37/ 65) vs 92.3％ (60/65);x2 =14.7,P＜0.01).Among 43 thyroid remnants and 22 metastatic lesions,99Tcm-MIBI imaging detected 39.5％ (17/43) of thyroid remnants and 90.9％ (20/22) of metastases,and those of 131I imaging were 100％ (43/43) and 77.3％ (17/22) respectively.The sensitivity of 131 I imaging in detecting thyroid remnants was significantly higher than that of 99Tcm-MIBI imaging(x2=24.0,P＜0.01).The sensitivities in detecting metastasis were not significantly different (x2=0.57,P＞0.05).The serum Tg level of positive groups (99Tcm-MIBI positive + 131I positive or 131I negative) were significantly higher than that of 99Tcm-MIBI negative + 131I negative group (t =-20.7 and-6.0,both P＜0.01),and that of 99Tcm-MIBI positive + 131I negative group was higher than that of 131I positive + 99Tcm-MIBI negative group(t=-2.7,P＜0.05).Conclusion 99Tcm-MIBI imaging could detect metastasis of DTC patient after first radioiodine therapy,but the value in detecting thyroid remnants is limited.

OBJECTIVE To investigate and compare the enzyme kinetic characters of psoralen (PRN)and isopsoralen(IPRN)in rat and human liver microsomes. METHODS PRN and IPRN in liver microsomes incubates were determined using LC-MS/MS. The enzyme kinetic and metabolic stability of PRN and IPRN were investigated by employing the optimized rat and human liver microsomes incubations. The Vmax and Km values were calculated using the nonlinear regression method. RESULTS The quanti?tative method showed good linearity within the range of 0.1-50.0 μmol · L-1 and was suitable for the assay in biological samples. The in vitro elimination was linear with the substrate concentrations lower than 1 μmol,the protein concentration within 0.5 g · L-1,and the incubation time within 40 min. The t1/2 values of PRN and IPRN in rat and human liver microsomes were 74.5,95.0,74.5 and 173.3 min, respectively. The Vmax values of PRN in rat and human liver microsomes were(1.140±0.080)μmol·min-1·g-1 protein,(0.620±0.060)μmol·min-1·g-1 protein,while Km values of PRN in rat and human liver microsomes were (12.9 ± 0.3)μmol · L- 1,(7.4 ± 1.3)μmol · L- 1,respectively. The Vmax values of IPRN in rat and human liver microsomes were(0.251±0.012)and(0.103±0.014)μmol·min-1·g-1 protein,while Km values of IPRN in rat and human liver microsomes were (3.0 ± 0.4)μmol · L-1,(3.4 ± 0.7)μmol · L-1,respectively. CONCLUSION The enzyme kinetic characters and metabolic stability of PRN and IPRN show species and chemical structures related differences. Interestingly,the metabolic eliminations of PRN and IPRN are similar in rats. However,the metabolic elimination of IPRN in humans involved in CYP enzymes may be much slower than that of PRN.

Objective To investigate the status quo of job-family and family-job conflicts among the nursing of the operating room nurses.Method A total of 190 nurses from an operating room participated in the survey by demographic questionnaire and work-family conflict scale.Results The average score on job-family was (3.15 ± 0.48) and there were significant differences between family-job conflict and job-family (2.75 ± 0.27 vs.3.55 ± 0.61) (t =-10.349,P＜0.05).There were significant differences in job-family conflict scores (P ＜0.05) in view of different age,working years,professional titles,number of night shifts per month,marital status and daily working hours.There were significant differences in family-job conflict subscale scores (P＜0.05).Conclusions The job-family conflicts of the nurses in the operating room is at the middle level.The perceived job-family conflict is higher than that of the family-job conflict.Such factors as age,length of work,professional tide,number of night shifts per month,marital status and daily working hours can lead to job-family conflict among the operating room nurses.The nursing administrative should displace nursing personnel and optimize the shifts so as to reduce the rate of job-family conflicts.