Objective:To observe effects of wrist-ankle acupuncture (WAA) onβ-endorphin (EP), nitric oxide (NO) in uterus tissue and prostaglandin F2α (PGF2α), substance P (SP) in serum of rats with primary dysmenorrhea. Methods:A total of 45 non-pregnant Wistar rats were randomly divided into a control group, a model group and a WAA group, 15 rats in each group. Rats in the model group and the WAA group received continuous abdominal subcutaneous injection of Diethylstilbestrol to establish dysmenorrhea rat models. On the first day after modeling, rats in the WAA group began to receive acupuncture on Point Lower 1 and Point Lower 2, once a day for 10 d. The control group and the model group didn’t receive any treatment. Writhing latencies and frequencies were recorded.β-EP and NO in uterus tissue homogenates and PGF2α, SP in serum were detected. Results:In the model group,β-EP and NO levels were the lowest among the groups, the serum PGF2α level was the highest, and serum SP level was the lowest. These measurements showed significantly difference between the model group and the control group (P<0.05). PGF2α in the WAA group was lower than that in the model group;β-EP, NO and SP levels were higher than those in the model group, with inter-group statistically significant differences (P<0.05). Conclusion: WAA may achieve analgesic effect through decreasing PGF2α, increasingβ-EP, NO and SP to relieve uterine cramps, increase blood flow and promote functional improvement.

Objective:To observe the effect of liquiritin on body weight and behavior in depression model rats.Methods:Seventy-two male Sprague-Dawley(SD)rats were randomly assigned into normal control group,model group(stress+vehicle),fluoxetine group(stress+ fluoxetine)and three liquiritin groups(stress+liquiritin at different dose).The behavior of rats was detected by sucrose consumption test and forced swimming test.Results:The model rats showed significantly lower body weight(429?45/494?37,P

Objective To explore the role of PI3K/AKT signaling pathway in total hepatic ischemia-reperfusion(I/R)-induced lung injury in rats.Methods This study was divided into 2 sub-projects.(1)36 rats were killed respectively at preischemia and after reperfusion,the lung tissue was then sampled.(2)12 rats were randomly divided into Wortmannin group and model control group. AKT,p-AKT protein expression,apoptotic cells and PCNA protein expression were tested respectively by Western blot,TUNEL and immunohistochemistry analysis.Results(1)Compared with that in preiscemia group,after I/R the apoptotic index (AI)was increased,the p-AKT/AKT ratio and PCNA-positive index showed bidirectional changes,and the histological changes were well identified.(2)p-AKT/AKT ratio showed a positive correlation with PCNA-positive index and a negative correlation with AI.(3)Compared with control group,the p-AKT/AKT ratio and PCNA-positive index were lower,histological changes was more significant and AI was higher in Wortmannin group. Conclusion PI3K/AKT signaling pathway had protective effect on lung injury induced by hepatic I/R,which was potentially mediated by anti-apoptosis and promoting proliferation.

Objective:To evaluate the uncertainty in carbamazepine ( CBZ) determination in human plasma by HPLC-MS/MS. Methods:The whole process of CBZ determination was analyzed and the uncertainty sources were established, and then the uncertainty was evaluated and combined, and the expanded uncertainty was also calculated. Results: The expanded uncertainty of CBZ with low (7.46 ng·ml-1) and high (745 ng·ml-1) levels was 0.410 ng·ml-1 and 33.400 ng·ml-1, respectively (P=95%, k=2). Conclusion:The uncertainty in CBZ determination in human plasma by HPLC-MS/MS is mainly caused by recovery, sample prepara-tion and matrix effect for low concentration, and by sample preparation and repeatability for high concentration.

OBJECTIVE:To establish and validate the method for the determination of carbamazepine (CBZ) in human plas-ma,and to apply the method for external quality assessment. METHODS:After precipitated with acetonitrile,the plasma sample was determined by LC-MS/MS. Using loratadine as internal standard,the determination was performed on Kromasil C18 column with mobile phase consisted of water (containing 0.1% formic acid)-acetonitrile (gradient elution) at flow rate of 0.6 ml/min and column temperature of 40 ℃. The ion transitions under MRM mode by ESI+ ionization were performed at m/z 237.1→194.0 and m/z 383.1→267.0 for CBZ and internal standard,respectively. RESULTS:The linear range of CBZ were 5-1 000 ng/ml (r>0.998). The limit of quantitation was 5 ng/ml. RSDs of inter-day and intra-day were 1.00%-6.42%;relative deviation were -6.93%-0.32%. The external quality assessment of 5 samples were 679.0,475.0,104.0,29.2 and 26.2 ng/ml,respectively. The pass rate of assess-ment result was 100%. CONCLUSIONS:The method is sensitive,accurate and specific. The method is applicable for the plasma concentration determination and external quality assessment of CBZ.

Objective:To evaluate the uncertainty in the determination of lamotrigine(LTG)in human plasma by LC-MS/ MS. Methods:The uncertainty sources in the determination of LTG were analyzed and the uncertainty was evaluated and combined. Re-sults:The expanded uncertainty of LTG at low(0. 050 4 μg· ml - 1 )and high(1. 27 μg· ml - 1 )concentrations was 0. 005 18 μg· ml - 1 and 0. 066 4 μg· ml - 1 ,respectively(P = 95% ,k = 2). Conclusion:The uncertainty in the determination of LTG in human plasma by LC-MS/ MS is mainly caused by the protein precipitation recovery,matrix effect and sample preparation at low concentration, and by the matrix effect,sample preparation and repeatability at high concentration.

Objective To study the relationship between expression of EGFR and E- cadherin and metastasis of esophageal cancer. Methods The expression of EGFR and E- cadherin in esophageal cancer tissues of 47 patients by LSAB immunohistochemistry method was studied. Results The positive rates of EGFR in distant metastasis group and local lymph node metastasis group were 47.62 % and 56.52 % respectively. They were higher than individual corresponding control groups(27.08 % and 26.32 %)(P

Objective: To observe the protective roll of atorvastatin on post-operative cardiac remodeling induced by transverse aortic constriction (TAC) in experimental mice with its possible mechanism.
Methods: A total of 48 C57BL/6 mice were randomly divided into 4 groups: Sham group, TAC group, TAC + valsartan group and TAC + atorvastatin group,n=12 in each group. Myocardial hypertrophy model was successfully established at 4 weeks after the operation, and then the animals were further treated by normal saline, valsartan 5mg/kg and atorvastatin 10mg/kg respectively for 8 weeks. Left ventricular anterior wall thickness at diastole (LVAWd), left ventricular posterior wall thickness at diastole (LVPWd), LVEF and left ventricular fractional shortening (FS) were examined by echocardiography, cardiac hypertrophy indexes were calculated. Protein expression of NF-κB was detected by Western blot analysis, cardiac tissue hydroxyproline (Hyp) level was measured by alkaline hydrolysis, serum levels of TNF-α, IL-1β were determined by ELISA, cardiac collagen deposition was identiifed by HE and Masson staining.
Results: Compared with Sham group, TAC group had increased LVAWd, LVPWd, cardiac hypertrophy indexes and increased area of cardiac fibrosis, allP<0.01; elevated protein expressions of NF-κB, Hyp, TNF-α, IL-1β, all P<0.01. Compared with TAC group, TAC + valsartan group and TAC + atorvastatin group presented improved cardiac hypertrophy indexes, decreased LVAWd, LVPWd and decreased area of cardiac ifbrosis, allP<0.01; reduced protein expressions of NF-κB, Hyp, TNF-α, IL-1β, allP<0.01.
Conclusion: Atorvastatin had protective roll on myocardial hypertrophy induced by pressure overload in experimental mice, which might be related to its anti-inlfammatory effect.

Objective To explore the effects of rosiglitazone (ROSI),a peroxisome proliferator-activated receptors-gamma (PPAR-γ) ligand,on hyperlipidemia in rats with severe acute pancreatitis (SAP) associated with lung injury.Methods A total of 120 male SD rats received intragastric administration of high fat diet for two weeks to induce experimental hyperlipemia.The hyperlipidemic rats were randomly (random number) divided into six groups:hyperlipidemia (HL) group (n =20),hyperlipidemia with SAP (HP) group (n =20),hyperlipidemia with rosiglitazone intervention (HRP) group (n =20),hyperlipidemia with rosiglitazone and antagonist to rosiglitazone (HRGP) group (n =20),rosiglitazone control (HR) group (n =20) and antagonist control (HG) group (n =20).The SAP was induced by a retrograde infusion of 5％ sodium tauroholate into bile-pancreatic duct,and the SAP was established in HP group,HRP group and HRGP group.In HL group,HR group and HG group,equivalent volume of normal saline was used instead of sodium taurocholate.In HRP group and HR group,ROSI (6 mg/kg) was administered via the femoral vein 1 hour prior to the administration of sodium taurocholate.In HRGP group,GW9662 (0.3 mg/kg),an antagonist to PPRA-gamma,was given via the femoral vein 30 min prior to the administration of ROSI.In HG group,only GW9662 (0.3 mg/kg) was given via the left femoral vein 30 min prior to pretend SAP modeling.Rats from each group were sacrificed by exsanguination 12 h after SAP modeling.Blood samples were taken from all subjects to measure serum amylase (AMY),total cholesterol (TC),triglycerides (TG),Successive sections of the paraffin embedded tissue from pancreas and lung were taken for pathological examination with hematoxylin-eosin (HE) staining.Histopathological changes of pancreatic and pulmonary tissues observed under light microscope were evaluated.In pulmonary tissue,nuclear factor-kappa B (NF-κB) p65 expression was assayed by immunohistochemistry.Intercellular adhesion molecule (ICAM-1) protein and tumor necrosis factor-α (TNF-γ) protein levels were studied using Western blot analysis.Results The serum levels of TC and TG in HL group and HP group were significantly higher than those in HR group and HRP group (1.24 ± 0.28,1.14 0.08 vs.0.41 ±0.17,0.58±0.12;14.86±1.47,12.42±0.96 vs.6.52±2.04,7.36±0.95,allP＜ 0.05);The levels of serum AMY,W/D ratio,pancreas pathologic score,lung pathologic score,expression of NF-κB p65,ICAM-1 and TNF-α in pancreas in the HP group and HRGP group were significantly higher than those in HL group,HR group,and HG group (6 501.9 ±3 770.0,5 922.2 ±925.9 vs.1 139.3 ± 35.6,1 070.8 ±67.0,1 012.4 ±94.7;3.14±0.16,3.06±0.12vs.1.81 ±0.13,1.76±0.23,1.83 ± 0.18;all P ＜0.05);Compared with the HP group and HRGP group,the levels of serum AMY,TC and TG were significantly decreased in HR group and HRP group,ameliorating pancreas and lung pathological damage,and down-regulating the expression of NF-κB p65,ICAM-1 and TNF-α in pulmonary tissue (all P ＜ 0.05).While there were no statistically significant differences in above biomarkers between HP group and HRGP group (all P ＞ 0.05).Conclusions Our study demonstrates that ROSI exerts anti-hyperlipidemic effect and anti-inflammatory effect on hyperlipidemia in rats with sodium taurocholate-induced severe acute pancreatitis associated with lung injury by inhibiting NF-κB and down-regulating the expression of TNF-α and ICAM-1.

OBJECTIVE:To evaluate the uncertainty of the determination of clopidogrel active metabolite derivative(CAMD) in human plasma by LC-MS/MS. METHODS:The course of CAMD determination in human plasma by LC-MS/MS were analyzed to evaluate the uncertainty caused by evaluate repeatability,weighing,standard solution preparation,biological sample prepara-tion,recovery,instrument precision and calibration curve fitting in CAMD assay;the uncertainty were calculated and expanded. RESULTS:The expanded uncertainty for low(0.107 ng/ml),medium(4.467 ng/ml)and high(155.667 ng/ml)levels of CAMD were 0.010 2 ng/ml,0.188 ng/ml and 6.413 ng/ml,respectively(k=2,P=95%). CONCLUSIONS:The uncertainty of the CAMD determination in human plasma by LC-MS/MS at low concentration is mainly caused by calibration curve fitting;at medium and high concentrations is mainly caused by biological sample preparation,extraction procedure,standard solution preparation and in-strument precision.