AIMTo study the protective effects of ramipril in combination with BQ-123 on myocardial ischemia/reperfusion (I/R) injury in vivo in anesthetized rats.

METHODSHealthy male Wistar rats were divided into 5 groups randomly and subjected to 30 min of myocardial ischemia followed by 120 min reperfusion. Ramipril, BQ-123 and their combination were given to rats respectively. To observe the protection of their combination against myocardial I/R injury. HR, MAP and the change of ST-segment were observed. Ventricular arrthymias were monitored. The activity of creatine kinase (CK) and lactate dehydrogenase (LDH) in plasma, the infarct size and morphologic change were examined.

RESULTSCompared with I/R group, the elevation of ST-segment was decreased. Onsets of VPC and VT were delayed, durations of VPC and VT were shortened, especially their combination. Incidences of VPC, VT and VF were decreased. Activity of plasma CK and LDH was decreased, especially their combination. IS, IS/AAR and the morphology of myocardium were improved, especially their combination.

CONCLUSIONRamipril, BQ-123 and combined using these two agents protected myocardium from I/R injury in vivo. The protective effects on delaying onset of VA, shortening duration of VA, decreasing the activities of CK and LDH, decreasing infrarct size and improving morphology of myocardium were better than using ramipril and BQ-123 alone.

Animals ; Arrhythmias, Cardiac ; prevention & control ; Cardiotonic Agents ; pharmacology ; Creatine Kinase ; blood ; Drug Synergism ; L-Lactate Dehydrogenase ; blood ; Male ; Myocardial Reperfusion Injury ; prevention & control ; Peptides, Cyclic ; pharmacology ; Ramipril ; pharmacology ; Rats ; Rats, Wistar

OBJECTIVETo study the role of serum 25-hydroxyvitamin D in the early diagnosis of vitamin D deficiency rickets.

METHODSConcentrations of serum 25(OH)D, calcium, phosphorus and alkaline phosphatase were measured in normal control (n=73), suspected rickets (n=45) and confirmed rickets groups (n=65). Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of serum 25(OH)D for rickets.

RESULTSSerum 25(OH)D levels in the suspected and confirmed rickets groups were 83±30 and 72±31 nmol/L respectively, which was lower than in the normal control group (112±37 nmol/L) (P<0.01). There was no significant difference between the suspected and confirmed rickets groups (P>0.05). Vitamin D deficiency rates in the suspected and confirmed rickets groups were higher than in the control group (P<0.01). The ROC curve area of serum 25(OH)D for the diagnosis of rickets was 0.760 (95%CI 0.692-0.820, P<0.01), and the optimal operating point was 90.70 nmol/L (sensitivity 68.49%, specificity 72.73%). There was no significant difference in levels of calcium, phosphorus and alkaline phosphatase between the three groups (P>0.05).

CONCLUSIONSSerum 25(OH)D levels in infants with suspected and confirmed rickets are significantly reduced and this may reflect vitamin D deficiency . Therefore, it may be useful to check serum 25(OH)D levels in screening for rickets.

Female ; Humans ; Infant ; Male ; ROC Curve ; Rickets ; blood ; diagnosis ; Vitamin D ; analogs & derivatives ; blood

Nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are the major enzymatic source of reactive oxygen species (ROS). NOX2 and NOX4 are expressed in the heart but its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion is unclear. This study investigated the effect of NOX on ANP secretion induced by hypoxia in isolated beating rat atria. The results showed that hypoxia significantly upregulated NOX4 but not NOX2 expression, which was completely abolished by endothelin-1 (ET-1) type A and B receptor antagonists BQ123 (0.3 µM) and BQ788 (0.3 µM). ET-1-upregulated NOX4 expression was also blocked by antagonists of secreted phospholipase A2 (sPLA2; varespladib, 5.0 µM) and cytosolic PLA2 (cPLA2; CAY10650, 120.0 nM), and ET-1-induced cPLA2 expression was inhibited by varespladib under normoxia. Moreover, hypoxia-increased ANP secretion was evidently attenuated by the NOX4 antagonist GLX351322 (35.0 µM) and inhibitor of ROS N-Acetyl-D-cysteine (NAC, 15.0 mM), and hypoxia-increased production of ROS was blocked by GLX351322. In addition, hypoxia markedly upregulated Src expression, which was blocked by ET receptors, NOX4, and ROS antagonists. ET-1-increased Src expression was also inhibited by NAC under normoxia. Furthermore, hypoxiaactivated extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) were completely abolished by Src inhibitor 1 (1.0 µM), and hypoxia-increased GATA4 was inhibited by the ERK1/2 and Akt antagonists PD98059 (10.0 µM) and LY294002 (10.0 µM), respectively. However, hypoxia-induced ANP secretion was substantially inhibited by Src inhibitor. These results indicate that NOX4/Src modulated by ET-1 regulates ANP secretion by activating ERK1/2 and Akt/GATA4 signaling in isolated beating rat hypoxic atria.

Nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are the major enzymatic source of reactive oxygen species (ROS). NOX2 and NOX4 are expressed in the heart but its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion is unclear. This study investigated the effect of NOX on ANP secretion induced by hypoxia in isolated beating rat atria. The results showed that hypoxia significantly upregulated NOX4 but not NOX2 expression, which was completely abolished by endothelin-1 (ET-1) type A and B receptor antagonists BQ123 (0.3 µM) and BQ788 (0.3 µM). ET-1-upregulated NOX4 expression was also blocked by antagonists of secreted phospholipase A2 (sPLA2; varespladib, 5.0 µM) and cytosolic PLA2 (cPLA2; CAY10650, 120.0 nM), and ET-1-induced cPLA2 expression was inhibited by varespladib under normoxia. Moreover, hypoxia-increased ANP secretion was evidently attenuated by the NOX4 antagonist GLX351322 (35.0 µM) and inhibitor of ROS N-Acetyl-D-cysteine (NAC, 15.0 mM), and hypoxia-increased production of ROS was blocked by GLX351322. In addition, hypoxia markedly upregulated Src expression, which was blocked by ET receptors, NOX4, and ROS antagonists. ET-1-increased Src expression was also inhibited by NAC under normoxia. Furthermore, hypoxiaactivated extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) were completely abolished by Src inhibitor 1 (1.0 µM), and hypoxia-increased GATA4 was inhibited by the ERK1/2 and Akt antagonists PD98059 (10.0 µM) and LY294002 (10.0 µM), respectively. However, hypoxia-induced ANP secretion was substantially inhibited by Src inhibitor. These results indicate that NOX4/Src modulated by ET-1 regulates ANP secretion by activating ERK1/2 and Akt/GATA4 signaling in isolated beating rat hypoxic atria.

Recombinant humanized anti-ricin monoclonal antibody (MIL50) is a recombinant humanized monoclonal antibody targeting ricin. In this study, an ELISA method was used to establish a method for the determination of MIL50 in macaque serum, and a cross design method was used. Twelve rhesus monkeys were intravenously injected 1 mg·kg^-1 test preparation (MIL50 freeze-died powder injection) and reference preparation (MIL50 liquid preparation) to determine the plasma concentration of MIL50 at different time points, and the pharmacokinetic parameters were analyzed to compare the pharmacokinetic characteristics of MIL50 liquid preparation and freeze-died powder injection in rhesus monkeys. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of the Chinese Academy of Medical Sciences and Use of Laboratory Animals and the regulations derived by the Animal Care and Welfare Committee of the Institute of Radiation Medicine, Academy of Military Medical Sciences (IACUC-DWZX-2020-503). The results showed that there was no significant difference between C_max and AUC_0-5d in the two groups. The liquid preparation was the reference preparation, with C_max ratio of 101.6% and AUC_0-5d ratio of 101.9%, the 90% confidence interval of C_max was 79.42%-129.92%, and the 90% confidence interval of AUC_0-5d was 85.72%-121.18%. These results suggested that different dosage forms of MIL50 had certain differences in the changes of blood drug concentration in rhesus monkeys.

Objective To make a better route for the patients needing routine or chemical treatment for long time with bad condition of peripheral veins by modifying the method of cathetering the internal jugular vein.Methods Thirty patients received modified method between 2008.01 and 2009.11. were selected as MJ group in this retrospective research. 87 patients were included for the usual cathetering into the internal jugular vein and selected as, Group J and 158 patients were as Group P for cathetering through the peripheral vein.According to the situation of catheters, indices, such as success ratio, staying duration and complication, were statistically coped with to analyze its advantage and disadvantage. Results There was no statistical difference of success ratio among three groups ( P > 0. 05 ), but the success ratio from Group P was up to 94.3% and was higher than those of the other two groups. The incidence of complication from Group MJ was 3.3%, lower than the other two groups ( P < 0. 05 ). The staying duration of Group MJ was obviously longer than the others.Conclusions The method of modified cathetering the internal jugular vein with PICC catheter did not limit the body. Its complications were shown less and its staying duration lasted longer. This method is a better route via deep vein and is worth spreading.

Objective To investigate metabolites of 1,5-dieaffeoylquin-ie acid (1,5-DCQA) in human urine after oral administration of 1, 5-DCQA. Methods Urine samples were collected 0-24 h after the oral administration of 600 mg dose of 1,5-DCQA, then the samples were pu-rified through C_(18) solid-phase extraction cartridge. The purified samples were analyzed by liquid chromatography and tandem electrospray ioniza-tion ion trap mass spectrometry (LC-ESI/MS/MS). Results A total of 28 metabolites were discovered, including methylated, glucuronidated and methyl-glucuronidated metabolites. Two important metabolites were determined by comparing with synthesized standards. Conclusion These results suggest that methylation, glucuronidation and isomerization are three important metabolic pathways of 1, 5-DCQA in human.

Healthy Beagle dogs were administrated with batroxobin by intravenous infusion at high, medium and low doses. The study of pharmacodynamics and pharmacokinetics was intended to clarify the relevance of them and provided strong evidence for clinical use of batroxobin. The blood samples were collected after injection based on the time schedule and samples were tested by ELISA method to get the concentration of batroxobin. At the same time, changes of prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT), fibrinogen (Fib) and D-dimmer were tested. The results showed that the concentration of D-D increased significantly after administration compared with that of before administration. The main pharmacokinetic parameters were as follows: t1/2 were (2.27 +/- 0.42) h, (10.65 +/- 2.19) h and (11.01 +/- 3.51) h; C(max) were (11.9 +/- 1.72) ng x mL(-1), (154.53 +/- 12.38) ng x mL(-1) and (172.14 +/- 47.33) ng x mL(-1); AUC(last) were (29.38 +/- 3.69) ng xh x mL(-1), (148.43 +/- 72.85) ng x h x mL(-1) and (599.22 +/- 359.61) ng x h x mL(-1). The elimination of batroxobin was found to be in accord with linear kinetics characteristics. The results of pharmacodynamics showed that D-dimmer level increased significantly after the administration of batroxobin, which was similar with the changes of batroxobin plasma concentration. Simultaneously, Fib concentrations in Beagle dog blood decreased significantly after the iv administration of batroxobin, while recovered to base level after 48 hours. PT, TT and APTT significantly became longer after administration, which returned to normal level after 48 hours. Especially, the D-dimmer levels and the batroxobin concentration in plasma after intravenous infusion of the drug were synchronized in Beagle dogs. Changes between PD/PK results had obvious correlation, and the D-dimmer levels in plasma can be one of the important monitoring indicators of batroxobin in thrombolytic medication.
Animals ; Area Under Curve ; Batroxobin ; administration & dosage ; blood ; pharmacokinetics ; pharmacology ; Dogs ; Enzyme-Linked Immunosorbent Assay ; Fibrin Fibrinogen Degradation Products ; metabolism ; Fibrinogen ; metabolism ; Fibrinolytic Agents ; administration & dosage ; blood ; pharmacokinetics ; pharmacology ; Infusions, Intravenous ; Male ; Partial Thromboplastin Time ; Prothrombin Time ; Thrombin Time

AIM: To investigate whether inactivation of extracellular signal-regulated kinase 1/2 ( Erk1/2 ) will affect the function of fibroblast growth factor 21 (FGF21) to regulate glucose and lipid metabolism .METHODS:Male db/db mice (8 weeks old) were treated with U0126 (an inhibitor of Erk1/2 kinase) for 1 week, and then treated with re-combinant human FGF21 protein and adenovirus-mediated FGF21 (Ad-FGF21).The profile changes of blood glucose and blood lipid were evaluated at 120 min or 4 weeks after FGF21 administration.Meanwhile, the molecular mechanism was ex-plored by in vitro study.RESULTS: Treatment of db/db mice with recombinant human FGF21 protein significantly re-duced blood glucose and triglyceride levels at 120 min after FGF21 administration , but these changes were comparable in U0126-treated mice .Furthermore , abnormal glucose and triglyceride levels , and glucose and insulin tolerance were strong-ly improved in db/db mice as accompanied with decreasing body fat content after 4 weeks of ad-FGF21 administration .In-terestingly, treatment with or without U0126 did not influence these effects of FGF21.Mechanically, treatment with Ad-FGF21 significantly upregulated the protein levels of p-Erk1/2 and peroxisome proliferator-activated receptor γ( PPARγ) as well as the expression of adiponectin at mRNA and protein levels in adipose tissues .However , treatment with or without U0126 did not change the profiles .On the other hand , in vitro experiments also indicated that treatment of adipocytes with recombinant human FGF 21 protein significantly activated Erk 1/2 phosphorylation , and upregulated the expression levels of PPARγand adiponectin (P<0.05).However, pre-administration of U0126 did not affect the profiles.CONCLUSION:Pharmaceutical inactivation of Erk 1/2 by U0216 does not affect the biological function of FGF 21 to regulate blood glucose balance and improve abnormal blood lipids in vivo.

OBJECTIVETo study the specific binding of the artificial clonal aryl hydrocarbon receptor translocator (ARNT) with the natural aryl hydrocarbon receptor (AhR) and the recolonization by polyclonal antibody. The dose-response relationship with tetrachlo-rodibenzo-dioxin (TCDD) was also studied to develop TCDD detection method and the binding degree related to dose response.

METHODS(1) The target genes including AhR-PAS, AhR-C and ARNT-PAS were amplified by RT-PCR by using the total RNA purified from the liver cells of C57BL/6J mice as templates to construct pGEX-5X1 recombinants. The recombinant plasmids were expressed in E. coli. (2) The rabbits were immuned by the clonal fusion proteins: AhR-PAS, AhR-C to prepare the polyclonal antibody. (3) The natural AhR from the hepatic cytosol of C57BL/6J mice was extracted. The artificial cloning expressed fusion protein:GST-ARNT-PAS and the natural AhR were incubated in different dose of TCDD. The quantity of the heterodimer through affinity adsorption and Western blots were measured.

RESULTS(1) The target proteins including AhR-PAS, AhR-C and ARNT-PAS were successfully cloned and expressed in E. coli. (2) The detection limit of polyclonal antibody AhR-PAS and AhR-C were 5 ng and 1 ng, respectively. (3) The total protein concentration prepared from the liver cells was 60.5 mg/ml. The artificial clonal protein ARNT-PAS could specifically bind to the natural AhR complex with the existence of TCDD. The detection limit of TCDD was 0.25 pmol which was 80 pg approximately.

CONCLUSIONA TCDD detection method based on the aryl hydrocarbon receptor system was established and the detection limit might reach pg grade.

Animals ; Cells, Cultured ; Limit of Detection ; Liver Extracts ; chemistry ; Mice ; Mice, Inbred C57BL ; Polychlorinated Dibenzodioxins ; analysis ; Rabbits ; Receptors, Aryl Hydrocarbon ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction