Objective To establish an HPLC method for the simultaneous determination of the 6 aconitum alkaloids in compoundTengwu Ointment.Methods Chromatographic column was Aglient XDB-C18 column (4.6 mm × 250 mm, 5μm), with the mobile phase A of acetonitrile- tetrahydrofuran (25∶8), phase B was 0.1 mol/L ammonium acetate solution (0.5 mL acetic acid added to the 1000 mL) with gradient elution;the flow rate was 1 mL/min;the detection wavelength was set at 235 nm;the column temperature was 25℃.Results Aconitine was in the good linear range of 0.072-0.648μg (r=0.999 5), with the average recovery of 99.29%, RSD=1.25%. Mesaconitine was in the good linear range of 0.062-0.648μg (r=0.999 5), with the average recovery of 99.12%, RSD=0.85%. Hypaconitine was in the good linear range of 0.064-0.384μg (r=0.999 8), with the average recovery of 99.57%, RSD=1.07%. Benzoylaconine was in the good linear range of 0.056-0.672μg (r=0.999 2), with the average recovery of 98.11%, RSD=0.61%. Benzoylmesaconine was in the good linear range of 0.055-0.993μg (r=0.999 9), with the average recovery of 99.27%, RSD=1.10%. Benzoylhypaconine was in the good linear range of 0.078-0.702μg (r=0.999 8), with the average recovery of 99.08%, RSD=1.38%.Conclusion This method is simple, sensitive, repeatable and accurate, which can be used for determination of aconitum alkaloids in compoundTengwu Ointment.

Objective To reveal the mechanism of Yuxuan granule(YXG) in treating pharyngitis from anti-inflammatory,expectorant,and antibacterial actions.Methods Xylene-induced ear edema and cotton pellet induced granuloma mice models were adopted to observe the anti-inflammatory effects of YXG Phenol red secretion in mice test was adopted to observe the expectorant activity of YXG Five bacteria by tube dilution method was adopted to observe antimicrobial action of YXG Besides,the changes of routine indexes in blood and tumor necrosis factor (TNF),interleukin-1 (IL-1) in serum were measured in acute pharyngitis models.Results YXG has shown a dose-dependent of its anti-inflammatory action.In the test of xylene induced mice ear swelling and cotton ball induced granuloma,the swelling degree of the control group,positive control groupand YXG high (6.14g/kg),medium (3.07g/kg) and low (1.53g/kg) concentration group were (9.00± 1.50) mg,(3.50±0.10)mg,(4.25±1.08)mg,(5.54±0.97)mg,(8.05±0.08)mg and (1.22±1.50)mg,(0.50±0.10)ag,(0.45± 0.08) mg,(0.75± 0.97) mg,(0.70 ± 0.08) mg respectively.YXG also had obviously expectorant effect to the golden staphylocoecus ameus,hemolytic streptococcus,streptococcus pneumoniae,salmonella typhi and corynebacterium diphtheriae,with the minimum inhibition concentration of 55,100,68 and 35 mg/ml respectively; YXG also reduced leukocyte count [percentage of leukocyte was (9.88± 1.22) × 109/L,(7.50± 1.58) ×109/L,(6.02± 1.05) × 109/L,(8.38± 1.13) × 109/L,(8.75 ± 1.08) × 109/L in the five groups respectively],neutrophils [the value was (35.62±5.00)×109/L,(28.49±3.40)×109/L,(28.89±5.22)× 109/L,(29.34±4.83)×109/L,(33.43 ± 4.27) × 109/L in the five groups respectively],lymphocytes [the value was (70.72± 5.04) ％,(67.89 ±4.37)％,(60.14±3.88)％,(65.38±4.45)％,(65.65±4.28)％ in the five groups respectively],TNF in blood serum [the value was (90.25±2.29)ng/ml,(80.23 ± 0.60) ng/ml,(76.83±0.33)ng/ml,(78.00 ± 0.34) ng/ml,(80.52±1.14)ng/ml in the five groups respectively] and IL-1 [the value was (222.06±50.22)pg/ml,(149.55±50.02)pg/ml,(130.37 ± 30.61)pg/ml,(158.68 ± 40.00)pg/ml,(178.76 ± 39.26)pg/ml in the five groups respectively] in the acute pharyngitis rat.Conclusion Through the anti-inflammatory,expectorant,antimicrobial activity and reducing levels of TNF,IL-1 YXG achieves the goal of treating pharyngitis.

Objective To optimize a preparation technique of Pingfenggujin Granule. Methods The technique was determined by the orthogonal design with content of Astragalosides, polysaccharides and the rate of dried extracts. The contents of polysaccharides and Astragalosides were determined by UV and ELSD-HPLC respectively. Results The optimum water-extraction technique was to steep 2.5 h, ten times of water as much as weigh of total drugs and extract three times, 2 h each. Conclusion The optimum technique is feasible, and the method of content determination is accurate and reliable.

AimTo study the anti-inflammatory effect of astragalosides(AST) and its mechanisms of action. Methods The exudate volume, neutrophil count and protein content in exudate were measuredinthe carrageenan-induced air pounch model in rats. The content of PGE2 was assayed by radioimmunoassay,the activity of PLA2 by microacid titration assay, IL-8 by ELISA, and NO by nitrate reductase assay. The production of O2 in neutrophil was determined by cytochrome C assay. Results AST(40, 80 mg· kg -1) could markedly reduce the exudate volume, neutrophil count, protein content, the content of IL-8, and the production of O2.AST lowered PLA2 activity of neutrophil and accellular component in exudate, and it also decreased the contents of PGE2 and NO in exudate. Conclusion AST has an obvious anti-inflammatory effect on carrageenan-induced acute inflammation.Its mechanisms may be related to the inhibition of vascular permeability and leukocyte migration, as well as to the suppression of PLA2 activity and the reduction of IL-8, PGE2, NO and O2 production.

Background Attack of primary acute angle-closure glaucoma (PAACG) often cause damage of retinal function.To measure the structural change after acute angle-closure glaucoma attack is of an important clinical significance.Flourier domain OCT(FD-OCT) is available.Objective This study was to measure and compare the thickness of peripapillary and macular retinal nerve fiber layer (RNFL) and macular volume in PAACG patients and normal subjects.Methods A case-controlled study was designed.Twenty-five eyes after acute angle-closure glaucoma attack and 25 fellow eyes were collected from April,2011 to February,2012 in Affiliated Eye Hospital of Wenzhou Medical College,and 25 eyes of normal subjects with the matched age and gender were enrolled at the same period.The thicknesses of RNFL and macula as well as the volumes of macula were measured by FD-OCT in all the eyes 2 weeks after acute angle-closure glaucoma attack.The outcomes were compared among the attacked eyes,fellow eyes and normal control eyes.Results The average thicknesses of RNFL were (125.72 ± 28.57),(108.36 ±9.31) and (106.10± 10.97)μm in the attacked eyes,fellow eyes and normal control eyes,showing a significant increase in the attacked eyes (all at P ＜ 0.05).The RNFL thickness values in the superior,interior and nasal quadrants at optical disc were significantly higher in the attacked eyes than those of the fellow eyes and normal control eyes (all at P＜0.05),but no significant differences were found in temporal and temporal-lower quadrants among the three groups (P=0.081,0.766).The thickness values of macular area were (283.72 ± 18.33),(280.28 ± 16.85) and (289.14±10.60)μm in the attacked eyes,fellow eyes and normal control eyes,with no significant difference among them (Fgroup =2.048,P =0.136),and fellow eyes were significantly reduced in comparison with the normal control group (P＜0.05),and those in the attacked eyes was not significantly reduced in comparison with the normal control group (P =0.224).The macular volumes were (5.589±0.355),(5.532±0.325) and (5.720±0.241)mm3in the attacked eyes,fellow eyes and normal eyes,without statistically significant difference among them (Fgup =1.027,P=0.363).The macular thickness values were lower in the outer race than those of the inner race and followed the pattern of nasal ＞superior＞ inferior＞temporal quadrants.At the temporal,superior,nasal and inferior quadrants of inner race,the temporal quadrant of the outer race,the central area,the macular thickness and volume values were declined in the attacked eyes and fellow eyes compared with the normal control eyes (all at P＜0.05).Conclusions The edema of RNFL is more serious than that of macula in the eyes within 2 weeks after acute angleclosure glaucoma attack.The retinas of macular zone are thinner,and the macular volumes are smaller in the attacked eyes and fellow eyes than those in the normal control eyes.

Objective To optimize the matrix formula of propranolol hydrochloride gels. Methods On the basis of single factor experiment, central composite design-response surface method was used to optimize the formula. Addition levels of glycerol (A), PEG-400 (B) and HPMC (C) were evaluated as the independent variables.Eight-hour accumulative penetration amount per unit area measured by HPLC was used as the index. Quadratic polynomial was used to estimate the relationship between the index and the independent variables, and to delineate response surface and overlay contour plots in order to select the optimal formulations.Finally, predicted responses were verified. Results The optimized formula consisted of 18.53% glycerol, 8.54% PEG-400 and 2.35% HPMC.The quadratic polynomial regression model of 8-h accumulative penetration amount per unit area:R1=-7 415. 69+306. 10A+167. 47B+4 820. 59C-8. 26A2-9. 81B2-1 025. 75C2 , and the cumulative transmittance was 49.6%. Conclusion A credible model is established by using central composite design-response surface method and the formula of propranolol hydrochloride gels is optimized and the gel is stable and controllable.

Objective To determine the eugenol in Dijin spray by GC-MS.Methods The chromato-graphic column was DB-5 fused silica capillary column.Eugenol was determined by program temperature risen method:the initial temperature was 60 ℃ for 1 min,then raised to 200 ℃ at the rate of 10 ℃/min and kept for 5 min,and at last raised to 250 ℃ at 15 ℃/min and kept for 5 min.Electron energy was 70ev.The ions of 164,149,and 77 were selected for eugenol detection.Results The calibration curve of eugenol was linear in the range of0.116～1.16 mg/ml (r=0.999 4) with the average recovery of 97.84％ (RSD=0.47％).Conclusion The method was quick,accurate and repeatable.It was suitable for the quality control of Dijin spray.

Objective To investigate the clinical effects of transurenthral resection of muscleinvasive bladder tumor (TURBT) in senile. Methods This study was clinical prospective random research.46 senile cases with muscle-invasive bladder cancer treated by TURBT was divided into 2 groups:control group and observed group.The observed group was treated with TURBT combined with intravesical instillation of pirarubi 30 mg immediately after TURBT,the bladder wall around tumor was resected thoroughly.The control group underwent radical cystectomy and pelvic lymph node dissection. Results The clinical improvement rate was higher in observed group than control group (85.7％ s.63.6％,X2 =5.3008,P=0.021).There was not difference in 3 years recurrence rate between observed group(25.0％) and control group(13.6％) (x2 =S.3008,P＞0.05).The 1 year survival rate were 95.8％,90.9％ and 3 years survival rate were 79.2％,72.7％ in observed group and control group,respectively,and there were no differences between the two groups(X2 =0.262,P=0.609). Conclusions TURBT combined with intravesical instillation of pirarubi can enhance clinical effect and reduce recurrence rate in the treatment of the elderly with muscle-invasive bladder cancer.

Objective To investigate the histogenesis,clinicopathological features and biological behavior of goblet cell carcinoid (GCC) of the appendix. Methods 3 cases of GCC of the appendix were studied by light microscopy and immunohistochemistry. Results 3 cases all occured in men. Microscopically: the tumor was composed of goblet cells containing mucin-filled with basal round to crescentic nuclei without atypia.These tumor cells were arranged in single forms,nests or acinus with no central lumen. 1 case was composed of simple goblet cells and typical carcinoid cells were seen in other 2 cases arranged in trabeculae and tubule.There was transition between goblet cells with tubelar and crypt epithelium.Immunohistochemical staining revealed that goblet cells were positively reacted with CgA, Syn, CEA, CK and p53 in 1 case. Conclusions Goblet cell carcinoid of the appendix arise from a pluripotent cell with divergent neuroendocrine and mucinous differentiation .It is a subtype of carcinoids of the appendix.The diagnosis mainly bases on its morphologic changes and immunohistochemical findings. The unpredictable behavior of this tumor is probably related to its component and the degree of infiltration.

Aim To study effects of loganin and morro-niside on aging astrocytes induced by D-galactose. Methods Cortex astrocytes of newly born rats were cultured in vivo and indentified by immunofluorescence method.Firstly,appropriate D-galactose concentration was selected and effects of loganin and morroniside on proliferation activity of aging astrocytes induced by D-galactose were determined by MTT test.Then SOD, MDA were taken as indicators to study the effects of loganin and morroniside on aging astrocytes induced by D-galactose.Thirdly,growth factors like gliar cell line-derived neurotrophic factor (GDNF),basic fibro-blast growth factor (bFGF)tested by ELISA method and expression of Bax,caspase-3,phospho-extracellu-lar signal-regulated kinese (p-ERK1 /2),phospho-mi-togen-actived protein kinase /extracellular signal-regu-lated kinase kinase (p-MEK1 /2)were taken as indi-cators to discuss the potential protection mechanism. Results Loganin and morroniside exerted certain effects on proliferation of aging astrocytes induced by D-galactose,improving SOD,GDNF,bFGF release and lowering MDA release significantly (P <0.05 ). The expressions of Bax and caspase-3 proteins had no difference between model group and loganin group, morroniside group.While the expressions of p-ERK1 /2,p-MEK1 /2 of loganin group,morroniside group were improved significantly compared with model group.Conclusion Loganin and morroniside have protective effects on aging astrocytes induced by D-ga-lactose,and increase the proliferation ability.Protec-ting their antioxidant systems and improving the expres-sion of p-ERK1 /2,p-MEK1 /2 proteins are possible mechanisms.