OBJECTIVE:To study the anti-oxidative effects of spirulina kinase (SPK) in vitro. METHODS:The methods of phenanthroine-Fe2+ oxidation method,DPPH and auto-oxidation of pyrogallol were used to measure the effects of different concen-trations of SPK on scavenging hydroxyl (OH-) free radical,DPPH free radical and superoxide anion (O2-) free radical;IC50 of SPK was calculated. Prussian blue reaction was used to determine total reducing ability(by absorbance)of different concentrations of SPK to Fe3+. Vitamin C(VC)was used as positive control in above trials. RESULTS:SPK could eliminate the OH-free radical, DPPH free radical and O2- free radical in concentration-dependant manner,and the maximum elimination rate of SPK to OH- free radical and DPPH free radical were 86.82% and 78.98%(IC50 were 54.31,0.636 g/L),which were higher than VC (64.77%, 73.49%). The maximum elimination rate of SPK to O2- free radical was 78.31%(IC50 was 3.918 g/L),which was lower than VC (94.14%). In reducing ability test,SPK improved absorbance in reducing ability test system,and maximum absorbance was simi-lar to VC in concentration-dependant manner. CONCLUSIONS:SPK has obvious anti-oxidant activities in vitro.

Objective : To understand the spatial distribution of multidrug resistant/rifampicin resistant pulmonary tuberculosis（MDR/RR PTB）in Wenzhou from 2014 to 2017,and to provide the scientific basis for MDR/RR TB control and prevention. Methods : The data of MDR/RR PTB cases in Wenzhou from 2014 to 2017 were collected from the Tuberculosis Management Information System of the Chinese Disease Prevention and Control Information System,and was associated with the geographic information database of Wenzhou Survey and Mapping Research Institute. The global spatial autocorrelation analysis was carried out by ArcGIS 10.1 to determine if there was spatial clustering of MDR/RR PTB cases in Wenzhou. The specific clustering areas of the MDR/RR PTB in Wenzhou was analyzed by SaTScan 9.3 and presented by ArcGIS. Results : There were 452 MDR/RR PTB cases reported in Wenzhou from 2014 to 2017,with a total registration rate of 4.74/100 000. The incidence rate of MDR/RR PTB in Wenzhou was unevenly distributed;the areas with registration rates of more than 7.45/100 000 were the north-central parts of Yongjia,the south-central parts of Yueqing and the east-central parts of Wencheng. The results of global spatial autocorrelation analysis showed that there were obvious clustering for MDR/RR PTB in Wenzhou（Moran's I=0.321,Z=7.352,P<0.001）. The spatial scanning found two clusters：20 towns/streets in the southeastern part of Yongjia and the south of Yueqing were the primary clustering areas（RR=2.213,LLR=22.353,P<0.001）;Yangyi Street and Shuangyu Street in Lucheng were the secondary clustering areas（RR=2.488,LLR=9.889,P=0.004）. Conclusion The MDR/RR PTB cases reported from 2014 to 2017 in Wenzhou had spatial clustering. The high-incidence areas were in the southeast of Yongjia,south of Yueqing,Yangyi Street and Shuangyu Street of Lucheng.

Objective In this study ,the effect of Spirulina kinase(SPK)on hemorheology ,platelet aggregation and cAMP in acute blood stasis rats model were investigated .Methods Forty SD rats were randomly divided into normal control group ,model group ,aspirin group and the high and low dose of SPK groups .All treatments were performed by gavage once a day for 7 days .Sub-cutaneous injection of adrenalin combined with ice water bath were used to establish the acute blood stasis rat model .The whole blood viscosity ,plasma viscosity ,red blood cell hematocrit (Hct) ,erythrocyte aggregation index (EAI) ,red blood cell deformation index(DI)anderythrocyterigidityindex(ERI)weredetectedbyautomaticbloodflow speedanalyzer .Meanwhile,thelevelofcAMP was detected by ELISA method .Rat platelet aggregation induced by ADP and the maximum aggregation rate was measured by tur-bidimetry .Results Results showed that SPK could significantly decrease the whole blood viscosity ,plasma viscosity ,Hct ,EAI and platelet aggregation rate ,increase the level of cAMP compared with model group(P<0 .05) ,but had no effect on DI and ERI .Con-clusion SPK can markedly improve the abnormal changes of hemorheology in acute blood stasis rats and inhibit the platelet aggregation .

OBJECTIVE:To study the anti-coagulation effect and mechanism of fibrinolytic enzyme SNFE in sipuculus nudus, and provide reference for further development of SNFE. METHODS:40 mice were randomly divided into blank control group(nor-mal saline),Xueshuantong group(positive control,15 mg/kg)and SNFE low-dose,high-dose group(15,30 mg/kg),10 in each group. After intravenous injection in tail,tail bleeding time (BT) and clotting time (CT) were respectively determined to investi-gate the anti-coagulation effect of SNFE. After taking blood in abdominal aorta of rats,test was divided into blank control group, positive control group and SNFE low-mass concentration,medium-mass concentration,high-mass concentration groups (0.25, 0.50,1.00 mg/mL). Prothrombin time(PT),re-calcium time(PRT)(using orokinase as positive drug,100000 U/mL),and max-mum platelet aggregation rate (PAG) in 5 min under adenosine diphosphate (ADP) inducer (using asprin as positive drug,0.50 mg/mL) were respectively determined,and anti-coagulation effect mechanism of SNFE was analyzed. RESULTS:Compared with blank control group,BT,CT of mice in each group were prolonged,with statistical significance in Xueshuantong group and SNFE high-dose group (P<0.05 or P<0.01). Plasma PT of rats in positive control group,SNFE medium-dose,high-dose groups and PRT in each administration group were significantly prolonged(P<0.05 or P<0.01);and PAG in administration group was signifi-cantly reduced(P<0.01). CONCLUSIONS:The fibrinolytic enzyme SNFE in sipuculus nudus can play its anti-coagulant effect by inhibiting the activity of coagulation factors in internal and external sources and ADP-induced platelet aggregation.

OBJECTIVE： To study the improvement effects of fibrinolytic enzyme from Sipunculus nudus （SNFE） on hemorheology disorder and vascular endothelium injury in naked acute blood stasis model rats. METHODS： SD rats were randomly divided into control group， model group， aspirin group （100 mg/kg） and SNFE high-dose and low-dose groups （2 500， 5 000     U/kg）， with 10 rats in each group. They were given relevant medicine intragastrically once a day， for consecutive 7 d. One hour after the 6th day of administration， except for control group， other groups were given adrenaline hydrochloride 0.8 mg/kg subcutaneously， and then the acute blood stasis model was induced by ice-water bath. Blood was collected from abdominal aorta 2 h after the next day. Blood rheological parameters such as whole blood viscosity （high， medium and low shear rate）， plasma viscosity， hematocrit， erythrocyte aggregation index and erythrocyte deformability index were measured by automatic rheometer. The contents of NO and ET-1 in plasma and their ratio were determined by ELISA， and the damaged degree of vascular endothelium were observed by HE staining. RESULTS： Compared with control group， whole blood viscosity of high， medium and low-shear rate， plasma viscosity， erythrocyte aggregation index and ET-1 content were increased significantly in model group， while erythrocyte deformability index， NO content and NO/ET-1 ratio were decreased significantly， with statistical significance （P＜0.05 or P＜0.01）. Compared with model group， whole blood viscosity of high， medium and low-shear rate， plasma viscosity， hematocrit， erythrocyte aggregation index and ET-1 content were decreased significantly in SNFE high-dose groups. Erythrocyte deformability index， NO content and NO/ET-1 ratio were increased significantly， with statistical significance （P＜0.05 or P＜0.01）. In SNFE low-dose group， erythrocyte deformability index and NO/ET-1 ratio were increased significantly， while ET-1 content was decreased significantly， with statistical significance （P＜0.05 or P＜0.01）. Vascular endothelial staining showed that compared with control group， the structure of aorta layers in model group was loose and disordered， the endothelial defect was incomplete， the vacuoles increased， and the endothelial damage was obvious. The endothelium of rats in each administration group was damaged to varying degrees， but the degree of injury was lighter than in model group. CONCLUSIONS： SNFE can improve hemorheological abnormalities and vascular endothelial injury in rats with acute blood stasis.

OBJECTIVE：To screen the effective compo nent in antioxi dant active fraction of Pueraria lobata . METHODS ：The antioxidant active fraction sample （S1-S20） of 20 batches of P. lobata were prepared. HPLC method was adopted. The determination was performed on SepaxBio-C 18 column with mobile phase consisted of methanol-water （gradient elution ）at the flow rate of 0.6 mL/min. The column temperature was set at 25 ℃，and detection wavelength was set at 250 nm. HPLC fingerprints of 20 batches of P. lobata were established by the Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 edition），and common peaks were identified. Cluster analysis ，principal component analysis （PCA）and orthogonal partial least squares discriminant analysis （OPLS-DA）were used to screen the effective components in antioxidant active fraction of P. lobata . RESULTS：There were 18 common peaks in HPLC fingerprints of 20 batches of antioxidant active fraction in P. lobata ，and the similarity was more than 0.99. Eight common peaks were identified ，which were 3′-hydroxypuerarin（peak 2），puerarin（peak 3）， 3′-methoxypuerarin（peak 4），daidzein（peak 5），genistein（peak 7），formononetin（peak 11），daidzein（peak 13）and genistein （peak 16）. The results of cluster analysis and PCA analysis showed that samples S 1，S3，S4，S6，S8，S18 and S 19 were clustered into one category ，and samples S 2，S5，S7，S9-S17 and S 20 were clustered into one category ；peak 2，peak 3，peak 10，peak 11 and peak 13 had great influence on principal component 1；peak 8 and peak 9 had great influence on principal component 2. OPLS-DA analysis showed that peak 4，peak 3，peak 2，peak 16，peak 13 and peak 11 had great influence on the quality of antioxidant active fraction of P. lobata . CONCLUSIONS ： HPLC fingerprint for active fraction of P. lobata is established in the study and 8 components are identified ；among them ， com puerarin，3′-hydroxypuerarin，daidzein and formononetin maybe the material basis of antioxidant fraction of P. lobata .