Aim To provide practical microassays for screening acetylcholinesterase (AChE) inhibitors in drug discovery.Methods The optimal conditions of assaying the activity of AChEs from electric eel,rat brain homogenate and cobra venom were determined in 96-well plates under physiological pH value and temperature by orthogonal matrix method.The concentrations of AChE,substrate and DTNB,and reaction time were optimized.After the effects of sample solvent (DMSO) used in the assay and stopping reagent on enzyme activity were assessed,the assay conditions were finally selected,and 492 kinds of extracts from Guizhou ethno-drugs were screened.Results Practical microassays for screening AChE inhibitors were successfully constituted by using AChEs mentioned above.The data analysis of screening results revealed that electric eel AChE possessed a high sentivity to inhibitors,and cobra venom AChE shared high similarity with rat brain homogenate in positive results.Conclusion Microassays constituted in this work possessed advantages of being easy,rapid,reliable,cost saving and flexible.AChE from electric eel was especially suitable for screening AChE inhibitors from extracts,and AChE from cobra venom was more suitable to be used in screening AChE inhibitors from large numbers of compounds.

Aim To provide practical microassay for screening butyrylcholinesterase(BChE) inhibitors in drug discovery.Methods The enzyme reaction was optimized in 96-well plates under physiological pH value and temperature by orthogonal matrix method.After the assay conditions were finally selected,3 inhibitors and 115 extracts from Guizhou ethno-drugs were tested.Results A practical microassay for screening BChE inhibitors was successfully constituted by using rat serum as the source of BChE.Conclusion The microassay constituted in this work possess advantages of being practical,rapid,reliable and economical.

Aim To provide practical microassay for screening ?-glucosidase inhibitors in drug discovery.Methods The optimal conditions of assaying the activity of ?-glucosidase were determined in 96-well plates under physiological pH value and temperature by orthogonal matrix method.Reaction time and the concentrations of ?-glucosidase and substrate were optimized.After the effects of sample solvent(DMSO) used in the assay and stopping reagent on enzyme activity were assessed,the assay conditions were finally selected.Then 492 kinds of extracts from Guizhou ethno-drugs were screened.Results Practical and sensitive microassay for screening ?-glucosidase inhibitors was successfully constituted.And in 492 kinds of extracts,145 kinds of samples effectively inhibited the enzyme activity.Conclusion The microassay constituted in this work possesses advantages of being rapid,sensitive,reliable,cost saving,easy and flexible.

A novel fibrinogenolytic protease,named atrase A,has been purified from the venom of Naja atra by sequential chromatography.Atrase A is a single chain glycoprotein with a molecular weight of 64.6 kD,an isoelectric point of pH 9.6 and a neutral sugar content of 4.16%.Atrase A specifically and slowly degraded α-chain of fibrinogen.This fibrinogenolytic activity Was inhibited by chelating agents(EDTA,EGTA and 1,10-phenanthroline)and DTY,and partially inhibited by PMSF,but not by soybean trypsin inhibitor,indicating it is a metalloproteinase.Atrase A showed edema-inducing activity and bactericidal activity against Staphylococcusa aureus.Atrase A did not show cytotoxicity on A549 and K562 cells in MTT assay,but detached adherent A549 cells from the substrate.Atrase A did not show significant inhibition of platelet aggregation induced by ADP or collagen,and did not exhibit proteolytic activities towards fibrin,azocasein and BAEE.It also did not show hemorrhage activity when injected subcutaneously into mice.

Objective To understand the dynamic status of schistosomiasis epidemic situation and Oncomelania hupensis snail status before and after the schistosomiasis transmission interrupted in the mountainous areas of Yunnan Province. Methods The data of schistosomiasis epidemic situation and snail status were collected and analyzed statistically in Jianchuan County from 10 years before the schistosomiasis transmission interrupted to 2008. Results The schistosomiasis control began in Jianch-uan County from 1954. In 1976,the criteria of schistosomiasis endemic controlled were reached,and the infection rate of popu-lation was 0.65%and the infection rate of snails was 0.40%. In 1981,the criteria of schistosomiasis transmission controlled were reached,and the infection rate of population was 0.34%and the infection rate of snails was 1.41%. In 1993,the criteria of schis-tosomiasis transmission interrupted were reached,and the infection rate of population was 0 and the infection rate of snails was 0. There was a fluctuation in the schistosomiasis epidemic situation and snail status during the whole control duration ,but the trend was decreasing. Conclusion The time from schistosomiasis endemic controlled to transmission controlled is relatively short,but the time from transmission controlled to transmission interrupted is relatively long. In the original schistosomiasis en-demic areas,there might be some areas where there is no the disease bud there still are snails.

Aim To study the development of acute lung inflammation in mice induced by activation of the complement alternative pathway and the changes of the related indicators, and to provide an ideal pathological model of acute lung inflammation in mice for drug screening and intervention. Methods Cobra venom factor( CVF) was used to activate complement alterna-tive pathway of SPF Kunming mice by intravenous injection. According to different sampling time, the mice were divided into 15 min, 30 min, 1 h, 2 h, 6 h group, and the parallel PBS control groups were set at the same time. Lung coefficient, lung water content, myeloperoxidase ( MPO ) activity, BALF cell number and protein content were tested. The pathological changes of lung tissue were observed by HE staining. The concentration of IL-6 , TNF-α, P-selectin and ICAM-1 in bronchoalveolar lavage fluid ( BALF ) and serum were determined by ELISA. Results CVF caused pulmonary inflammatory cell infiltration in mice obviously. Compared with PBS groups, MPO activity of lung tissue, BALF cell and the protein concentration were significantly increased. The contents of IL-6, TNF-α, P-selectin in BALF and serum were in-creased, and the content of ICAM-1 in serum was also increased. The content of P-selectin in BALF reached the first peak at 30 min point, the content of IL-6 and TNF-α in BALF reached the first peak at 1 h point, but the indicators had no further changes at 2 h point, and all the indicators rose again at 6 h point. The lev-els of IL-6 and TNF-α in serum reached peak at 1 h point,then the content showed lower levels at the sub-sequent time points. The levels of P-selectin and ICAM-1 in serum increased along the time. Lung coef-ficient, lung water content and ICAM-1 of the BALF showed no significant alteration. Conclusion The ac-tivation of the complement alternative pathway can lead to acute lung inflammation in mice and the inflammato-ry response is the most obvious at 30 min to 1 h. The study could provide an ideal pathological model of a-cute lung inflammation in mice for drug screening and intervention.

Objective To investigate the inflammation levels of 2-diabetes patients before and after 3 months of improving glycemic control.Methods A longitudinal study was performed in a subgroup of 48 subjects with T2D and poor glycemic control.Forty-four healthy individuals were taken as control group.The serum concentration of C-reactionprotein (CRP),interleukin-6 (IL-6),interleukin-6 (IL-8),transforming growth factor-β1 (TGF-β1) and transforming growth factor-β1 (MCP1) in all participants were measured simultaneously by multiplexed Luminex assay.Results The serum levels of CRP,MCP-1 of 2-diabetes patients were 3.96 (3.45,5.58) mg/L and (195.0± 129.8) ng/L,significant higher than those in control group (2.25 (1.24,3.22) mg/L,(148.5±85.7) ng/L),and the differences were significant(t=-2.580,P=0.010;t=-2.118,P =0.047).No significant difference was found in the serum levels of IL-6,IL-8,TGF-β lbetween the two groups (P＞0.05).TGF-β1 level in patients with good glycemic control decreased to 26.85 (23.17-31.12) ng/l,significant lower than that before glycemic control (43.5(26.5-62.25) g/L;Z=-2.191,P=0.028),and there were no significant differences among the other 4 kinds of inflammatory factors before and after blood glucose control(CRP:Z =-0.937P =0.372;IL-6:Z =-0.875,P =0.396;IL-8:Z =-1.215,P =0.286;MCP-1:t =-1.846,P=0.065).Conclusion Low grade systemic inflammation status in T2D patients.Improvement of glycemic control reduces TGF-β1 levels and plays a key role in delaying the development of diabetic nephropathy.

Objective To determine the baseline echocardiographic characteristics and the time course and degree of recovery of left ventricular (LV) systolic dysfunction in patients with tachycardia-induced cardiomyopathy ( TCM ) . Methods Seven hundred and fifteen patients received radiofrequency cather ablation ( RFCA ) for tachycardiarrhymias from July 2010 to July 2013 were screened in this study. Only 33 patients with reduced left ventricular ejection fraction (LVEF) (LVEF<50% and improved≥15%) were diagnosed with tachycardia-induced cardiomyopathy and were included in the study. Patients with early improvement (over 25%increase in LVEF at 1-week follow-up compared to the baseline ) were enrolled in the improved group , and the rest patients were enrolled in the improved group. All Patients received transthoracic echocardiography for LV size and function detection at 1 week and at 3,6,12 months follow-up. Results The average baseline of the LV end-diastolic diameter, and the LVEF were (55 ± 10.7)mm and (38 ± 4.6)%, repectively. Early improvement ( over 25%increase in LVEF at 1-week follow-up compared to the baseline ) in the improved group was observed in 16 patients. Patients with early improvement had higher LVEF at 12-month follow-up compared to the patients without early improvement [(69.2± 4.2)% vs (58.1 ± 6.9)%, P < 0.001]. Conclusions RFCA is proved to be a relatively safe and effecient treatment method. Atrial fibrillation related to TCM , rhythm control is superior to the rate control. The early improvement in LVEF may potentially predict the complete reversibility of LV systolic dysfunction.

Objective To explore the rescue and nursing of complicated acute cardiac tam-ponade (ACT)in patients with cardiac interventional operation (CIO).Methods A total of 64 CIO patients with complicated ACT were randomly divided into observation group and control group,32 cases in each group.Both groups were conducted with emergency treatment for ACT, and on this basis the control group was treated with routine nursing,while the observation group was treated with comprehensive nursing.Discovery rate of ACT,success rate of rescue as well as satisfactory degree toward nursing and HDL score were compared between two groups.Results In the observation group,the discovery rate of ACT and success rate of rescue were significantly higher than those in the control group (P <0.05).Satisfactory degree toward nursing in the ob-servation group was significantly better than that in the control group (P <0.05).After nursing, scores of anxiety and depression as well as total score of HDL in the observation group were signifi-cantly lower than those before treatment and the control group (P <0.05).Conclusion For CIO patients with complicated ACT,rescue measures and comprehensive nursing can significantly im-prove the success rate of rescue and satisfactory degree toward nursing,reduce the HDL score and reduce medical disputes.

Objective To explore the rescue and nursing of complicated acute cardiac tam-ponade (ACT)in patients with cardiac interventional operation (CIO).Methods A total of 64 CIO patients with complicated ACT were randomly divided into observation group and control group,32 cases in each group.Both groups were conducted with emergency treatment for ACT, and on this basis the control group was treated with routine nursing,while the observation group was treated with comprehensive nursing.Discovery rate of ACT,success rate of rescue as well as satisfactory degree toward nursing and HDL score were compared between two groups.Results In the observation group,the discovery rate of ACT and success rate of rescue were significantly higher than those in the control group (P <0.05).Satisfactory degree toward nursing in the ob-servation group was significantly better than that in the control group (P <0.05).After nursing, scores of anxiety and depression as well as total score of HDL in the observation group were signifi-cantly lower than those before treatment and the control group (P <0.05).Conclusion For CIO patients with complicated ACT,rescue measures and comprehensive nursing can significantly im-prove the success rate of rescue and satisfactory degree toward nursing,reduce the HDL score and reduce medical disputes.