Objective To observe the protection of Caspase inhibitor(zVAD-fmk,benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone) on neonatal rat with hypoxic-ischemic brain damage(HIBD).Methods Thirty-six neonatal rats,7 days old,were randomly divided into hypoxic-ischemic(HI) control group(A),zVAD-fmk treated group(B) and sham group(C).Before HI insult,a pan-Caspase inhibitor,zVAD-fmk or normal buffer solution was injected into the cerebral ventricle.The water content of cerebral hemisphere was measured and the percentage of apoptofic cells in hippocampal neurons was measured by Flow cytometer(Annexin V/PI) at 24 hours after HI insult.The effect on body weights(percentage of increased weight,WIP) and macroscopical changes(percentage of cortox and hippocampal dead neurons) were assessed at 14 days.Results Compared with group A,the water content of ischemic hemisphere and apoptosis percentage of hippocampal neurons in group B reduced significantly.The difference of percentage of increased weight at 14 days in group B was not significantly.Microscopic examination showed that cortox and hippocampus neural death rate in group B was proved significantly reduced compared with that in group A.Conclusion Intracerebral administration of zVAD-fmk has protective effects on hypoxic-ischemic brain damage in neonatal rat.

Objective To investigate the relation between serum adiponectin and coronary artery calcification score (CACS), and find the risk factors for CACS in patients with chronic kidney disease (CKD). Methods Twenty-nine patients with 3-5 stage CKD were selected. The serum adiponectin was measured by enzyme linked immunosorbent assay. The heart was scanned by 64-row spiral CT, and the CACS was calculated. The blood calcium, phosphorus, intact parathyroid hormone, total cholesterol, low density lipoprotein cholesterol, albumin, urea nitrogen, creatinine, uric acid and high sensitive C reactive protein levels were measured, and the calcium-phosphorus product and estimation glomerular filtration rate were calculated. Results In 29 patients with CKD, 24 cases (83%) had coronary artery calcification with different degree (CACS>0 score), and the average CACS was 508 (0-3 363) scores. There were statistical differences in systolic blood pressure, urea nitrogen and estimation glomerular filtration rate between CKD patients with CACS≥100 scores (15 cases) and CKD patients with CACS<100 scores (14 cases):(146.00± 13.00) mmHg (1 mmHg=0.133 kPa) vs. (123.00±9.00) mmHg, (15.44±8.36) mmol/L vs. (9.71±2.52) mmol/L and (21.77 ±11.81) ml/ (min·1.73 m2) vs. (38.71 ±11.56) ml/ (min·1.73 m2), P<0.01 or <0.05. Pearson correlation analysis results showed that CACS had positive correlation with systolic blood pressure, creatinine and uric acid, and negative correlation with albumin and estimation glomerular filtration rate. Mult-stepwise regression analysis results showed that systolic blood pressure and estimation glomerular filtration rate were the independent risk factors of CACS. Conclusions The patients with 3-5 stage CKD have severe coronary artery calcification. The systolic blood pressure and estimation glomerular filtration rate are the independent risk factors of coronary artery calcification.

OBJECTIVETo explore action mechanism of electroacupuncture (EA) at "Fenglong" (ST 40) for treatment of hyperlipidemia.

METHODSForty SPF-grade SD rats were randomly divided into a normal group (group A), a model group (group B), a model diet-control group (group C), a model + EA group (group D) and model diet-control + EA group (group E), 8 cases in each one. The rats in group A were fed with normal diet continuously while those in the rest 4 groups were fed with high-fat diet to establish hyperlipidemia model. Afterwards, the rats in group C and group E were fed with normal diet, while EA at "Fenglong" (ST 40) was applied in group D and group E, 30 min per time, once a day. After 28 days of treatment, macrophage was collected in each group. Oil red O-staining was applied to detect the formation of foam cells, and enzyme-linked immunosorbent assay (ELISE) was adopted to measure the contents of total cholesterol (TC) in macrophage and analyze the rate of cholesterol efflux.

RESULTSThe counts of positive cells of oil red O-staining and the contents of TC in the group B, group C and group D were significantly increased compared with those in the group A (all P < 0.01). The counts of positive cells and contents of TC in the group C, group D and group E were significantly clecreased compared with those in the group B (all P < 0.01), and the decline in group D was more obvious than that in the group C (all P < 0.01). Compared with group C and group D, the counts of positive cells and contents of TC in the group E was obviously decreased (all P < 0.01), which was not statistically different from group A (P > 0.05). In the meantime, compared with group C, the rate of cholesterol efflux in group D and group E was significantly increased (both P < 0.01), and the rise in group E was more obvious than that in the group D.

CONCLUSIONThe electroacupuncture at "Fenglong" (ST 40) could significantly prohibit the transformation of macrophage into foam cell and increase rate of cholesterol efflux in macrophage, which could prevent and reverse the formation of foam cell and play an essential role in treating hyperlipidemia and stopping it from developing into a further level.

Acupuncture Points ; Animals ; Biological Transport ; Cells, Cultured ; Cholesterol ; metabolism ; Electroacupuncture ; Foam Cells ; metabolism ; Humans ; Hyperlipidemias ; metabolism ; therapy ; Macrophages ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Objective:To study the pharmacokinetics of propranolol in Wistar rats after acute exposure to high altitude.
Methods:Fourteen male Wistar rats (200±20) g were selected. After administration of propranolol tablets (0.05 g/kg, i.g.), blood samples (3 mL) were collected at 0, 20, 40 min,1, 1.5, 2, 4, 6, 8, 12 and 24 h, respectively. The pharmacokinetic parameters were determined by LC-MS/MS and DAS 2.0 software.
Results:The main pharmacokinetic area under concentration-time curve (AUC), mean retention time (MRT), half-life (t1/2) and peak plasma concentration (Cmax) of propranolol were increased by 442.61%, 47.45%, 73.13%and 352.97%, respectively, whereas Tmax and clearance (CL) were decreased by 80.87%and 68.94%, respectively.
Conclusion:This study displays significant changes in the pharmacokinetics of propranolol under high altitude, which may provide evidence for clinical rational application of propranolol at high altitude.

Objective To study the effects of imatinib mesylate on the apoptosis in human melanoma cell line M14.Methods M14 cells were cultured in vitro in the presence of imatinib mesylate at three concentrations(5,10 and 20 μmol/L)for 96 hours.Sebsequeutly,annexin V-FITC and propidium iodide(PI)double staining flow cytometry and terminal deoxvnucleotidvl transferase (TdT)-mediated dUTP nick end labeling (TUNEL)were used to detect the cell cycle and apoptosis,respectively,DAPI staining to observe the mot-phological changes.and Western blot to measure the protein expressions of bcl-2 and bax in cells.Results Imatinib mesylate of the three concentrations could induce an evident increase in the apoptosis in M14 cells.Compared with untreated M14 cells,an increase of cell population in S phase was observed in imatinib mesylate.treated cells(P<0.05),along with a decline in cell population in G2/M phases(P<0.01).Annexin V/PI double staining and TUNEL revealed a significant increase in the rate of early apoptosis and in the acount of apoptotic cells,respectively,in M14 cells treated with imatinib mesylate of the three concentrations(all P<0.01).After treated with imatinib mesylate of 20 μmol/L.there was a morphological change characteristic of apoptosis in M14 cells,together with an upregulated expression of bcl-2(t=15.46,P<0.01)and downregu-lated expression of bax(t=25.53,P<0.01).Conclusions Imatinib mesylate can interfere with the process of cell cycle of and induce the apoptosis in M14 cells,which may be mediated through mitochondrial pathway.

Objective:To study the effects and safety of metformin combined with vildagliptin on the glycemic control for patients with newly diagnosed type 2 diabetes mellitus.Methods:60 patients with type 2 diabetes mellitus who were treated from February 2015 to April 2016 were selected and divided into the control group and the observation group according to different treatment methods.The control group was treated with routine treatment.The observation group was treated with vildagliptin based on the control group.The blood glucose,glycosylated hemoglobin,two-hour postprandial blood glucose and serum as well as urinal amylase were measured before and after treatment,and the clinical curative effect of the two groups and the levels of interleukin-6,tumor necrosis factor and C-reactive protein were compared.Results:After treatment,the total effective rate of observation group was 90％,which was significantly higher than that of the control group(66.7％,P＜0.05).After treatment,the serum interleukin-6,tumor necrosis factor,C-reactive protein and fasting blood glucose,glycosylated hemoglobin and postprandial blood glucose levels were significantly lower than those of the control group[(7.63± 1.12)dvs(8.68± 1.30)d;(7.23± 0.95)d vs(7.89± 1.20)d;(11.14± 1.56)d vs(12.12± 1.89)d];[(12.12± 1.89)d vs(ll.20± 1.34)d;(6.89± 0.96)d vs(8.23± 1.10)d;(1.65± 0.23)d vs(3.65± 0.48)d] (P＜0.05).After treatment,the INS level of observation group was significantly lower than that of the control group (P＜0.05) and the GLP-1 level was significantly higher than that of the control group (P＜0.05).Conclusion:Metformin combined with vildagliptin could effectively control the blood glucose of patients with newly diagnosed type 2 diabetes and enhance the safety.

Membrane proteins are the main undertakers of biofilm function, and also the most important target group for innovative drug discovery and research. About 60% of drugs targets are membrane proteins. Due to the obvious aggregation and denaturation tendency of membrane proteins in aqueous solution, it is difficult to simulate the membrane like environment to maintain the correct conformation of membrane proteins in vitro, which results in the slower-growing research on the structure and function of membrane proteins and related ligand drugs than that of water-soluble proteins. Membrane protein stabilization technology is the premise of establishing high specificity, high sensitivity and high throughput drug screening methods for membrane protein ligands, which is of great significance. In this paper, some techniques for stable separation and purification of membrane proteins are reviewed, including detergents, artificial membranes, polymers, lentiviral particles and so on, as well as their specific applications in drug screening.

Objective To explore the effect of application of hospital-school combined three-dimensional teaching mode in the operation nursing practice teaching.Methods 129 students were collected by cluster sampling,then based on class unit,they were randomized into the experimental group (69 students) and the control group (60 students).The experimental group used hospital-school combined threedimensional teaching mode and the control group used traditional teaching mode.Then,both groups had finished an operation nursing comprehensive theoretical examination and a self-report questionnaire for mastery of basic knowledge of operation nursing.Besides,the experimental group had a questionnaire survey of cognitive evaluation for the new teaching mode.Results The average score of operation nursing comprehensive theoretical examination of the experimental group was (77.81 ±9.87),higher than the control group (70.35±12.37); The self-report score for mastery of basic knowledge of operation nursing of the experimental group was (3.94±0.45),higher than the control group (3.68±0.46).The students of the experimental group also considered that the new teaching mode improved their comprehensive knowledge and innovative abilities,critical thinking ability,humanistic care spirit,communication ability,team cooperation ability and innovation ability,etc.Conclusions The effect of application of hospital-school combined three-dimensional teaching mode in the operation nursing practice teaching was good,but it still had some limitations which needed us to pay attention to and to perfect.

Objective To explore the changes of high-sensitivity C reactive protein(hs-CRP) and platelet parameter levels in newborns with hypoxic-ischemic encephalopathy(HIE)and its clinical significance.Methods Thirty-five infants with HIE and 16 healthy newborns were selected as study cases and controls respectively by automantc biochemistry analyzer.Serum hs-CRP content was measured by reaction rate method;Platelet parameter levels were detected by collecting blood samples from peripheral vascular of heel,and the activity of creatine kinase(CK),creatine phosphokinase isoenzyme(CK-MB),lactate dehydrogenase(LDH),alpha-hydroxybntyric dehydrogenase(?-HBDH),alanine transaminase(ALT),aspartate aminotransferase(AST) were assayed as well.Results 1.The hs-CRP levels in newborns with HIE increased obviously in acute stage,and significant difference were observed compared with controls(P0.05).2.PLT in newborns with HIE decreased significantly in acute stage compared with that of controls(P0.05).3.The values of CK,CK-MB,LDH,?-HBDH,AST,ALT in newborns with HIE were significantly higher than those in controls in acute stage(P

Liquid-phase microextraction is a novel pretreatment technique for biological samples developed on the basis of liquid-phase extraction technology, which is simple, rapid, economical, and environmentally friendly, and has been widely used in the analysis of biological matrix samples such as blood, urine, and saliva. In this paper, we review the basic principles of the main modes of liquid-phase microextraction techniques, i.e., single-drop microextraction, dispersive liquid-liquid microextraction, and hollow-fiber liquid-phase microextraction, and the progress of their applications in biological sample pretreatment by reviewing the literature in the past five years, with a view to providing technical support and reference for sample pretreatment in the fields of in vivo drug analysis, pharmacokinetic studies and new drug development.