OBJECTIVE:To optimize the extraction process for Renzhi anti-cough granules.METHODS:The preparation process was optimized by orthogonal experiment design L9(33) with the amount of solvent,extraction times and concentration of alcohol as factors and with yield of paste,the latency for the cough of mice and the cough times as indexes. RESULTS:The optimum extraction process was as follows:Adding 5 times of solvent for extraction twice,and the extractum obtained was dissolved with 60% alcohol and stayed still overnight,then the supernatant was taken for filtration with ethanol-extract filtrate recovered under reduced pressure until paste was obtained,then 4 times of thoroughly mixed mixture of sugar-starch (3∶2;the starch was dried at 105 ℃ for 1 hour in advance) was added to prepare soft material,after sieving,drying and granulation,porous yellowish granules were obtained.CONCLUSION: The method can help reduce the dosage needed while enhancing drug efficacy and it is applicable for the preparation of Renzhi anti-cough granules.

OBJECTIVE:To provide reference for rational use of proton pump inhibitor(PPI)in the clinic. METHODS:In ret-rospective study,the utilization of PPI in Zhangye Municipal People’s Hospital during 2012-2014 were analyzed statistically in re-spects of consumption sum,DDDs,DDC,etc. RESULTS:The consumption sum and DDDs of PPI were increasing year by year, pantoprazole and omeprazole occupied the first and the second place in the list of consumption sum during 2012-2014. CONCU-SIONS:The consumption sum of PPI is well synchronous with their DDDs,and the application of proton pump inhibitors is reason-able basically.

OBJECTIVE:To study the improvement of Zea mays extract on cholestatic liver diseases.METHODS: Acute cholestatic liver injury model of rat was induced by bile duct ligation and cholestatic liver model of rats was established by ANIT.Model rats were given 6.2 g?kg-1 and 3.1 g?kg-1 dose of Z.mays extract for 5 days.The content of TBIL,DBIL,ALT,AST and TBA in serum and the secretion of bile were determined.RESULTS: The corn stigma extract decreased the content of DBIL,ALT and TBA in serum of ANIT-inducing cholestatic liver model,and increased the flow and speed of bile (P

BACKGROUND: Islet transplantation is an effective method for the treatment of type 1 diabetes mellitus and parts of type 2 diabetes mellitus. However, its application is hindered by insufficient sources and immunologic rejection. Though transdifferentiation of pancreatic stem cells is at the starting step, it is thought to be the hopeful source for islet cell transplantation.OBJECTIVE: To look for a suitable cells-transplantation source for the treatment of diabetes mellitus. METHODS: The pancreatic ductal epithelial cells were separated from Kunming mice and cultured in DMEM/F12 medium supplemented with keratinocyte growth factor, hepatocyte growth factor and nicotinamide, etc. Samples were taken at different time points for light microscopy and electron microscope. The changes of CK-19 and PDX-1 were detected by immunocytochemistry at 1 and 16 days. The expressions of insulin and glucagon gene were detected by RT-PCR at 1 and 16 days. The physiologic function of these islet-like clusters was determined by dithizone staining and glucose stimulation at 21 days. RESULTS AND CONCLUSION: A large number of epitheliod cells were CK-19 immunoreactive positive and few of them were PDX-1 positive at 1 day after isolation, then CK-19 positive cells proliferated quickly and formed substantial plaques of epithelial cells in cobblestone pattern. At 16 days later, these cells begin to form islet-like clusters gradually, while most of them were PDX-1 immunoreactive positive. The analysis of mRNA by RT-PCR showed very low levels of insulin and glucagon mRNA in the starting materials but increase was found as the process of transdifferentiation. At 21 day differentiated islet-like clusters were stained red by dithizone. In those samples exposed to a stimulatory 15 mmol/L glucose, there was a 1.6-fold increase in insulin compared with to 5.6 mmol/L glucose (P < 0.05). Pancreatic ductal cells of adult Kunming mice could proliferate quickly and have the potency of transdifferentiation into islet-like clusters when cultured in vitro under appropriate conditions.

A double targets of high throughput screening model for xanthine oxidase inhibitors and superoxide anion scavengers was established. In the reaction system of xanthine oxidase, WST-1 works as the probe for the ultra oxygen anion generation, and product uric acid works as xanthine oxidase activity indicator. By using SpectraMax M5 continuous spectrum enzyme sign reflectoscope reflector, the changes of these indicators' concentration were observed and the influence factors of this reaction system to establish the high throughput screening model were studied. And the model is confirmed by positive drugs. In the reaction system, the final volume of reaction system is 50 μL and the concentrations of xanthine oxidase is 4 mU x mL(-1), xanthine 250 μmol x L(-1) and WST-1 100 μmol x L(-1), separately. The Z'-factor of model for xanthine oxidase inhibitors is 0.537 4, S/N is 47.519 9; the Z'-factor of model for superoxide anion scavengers is 0.507 4, S/N is 5.388 9. This model for xanthine oxidase inhibitors and superoxide anion scavengers has more common characteristics of the good stability, the fewer reagent types and quantity, the good repeatability, and so on. And it can be widely applied in high-throughput screening research.

Objective To assay the early quality of life and posttraumatic stress disorder (PTSD) and relating influential factors in patients with severe blunt chest trauma (sBCT).Methods Demographic and clinical data of sBCT patients treated between January 2011 and December 2011 were collected.Early quality of life and PTSD symptom level at posttraumatic months 1,3,and 6 were analyzed by using short form 36 health survey (SF-36) and impact of event scale-revised (IES-R) respectively.Furthermore,logistic regression analysis was performed to identify the risk factors associated with quality of life of the patients.Results A total of 107 patients were included in the study.Ultimately,83 patients were available to the 6-month follow-up.A low score for SF-36 remained at posttraumatic 6 months and one-third of the 83 patients sustained mild or severe PTSD symptoms.Major influential factors to posttraumatic quality of life included age,ISS ≥ 20,combined craniocerebral injury,combined spinal and pelvic injuries,posttraumatic complications,and PTSD.Conclusions Early quality of life in sBCT patients is poor.Therefore,the early intervention with identification of specific risk factors is contributive to better quality of life.

OBJECTIVE To investigate the blocking activities of a series of potential α1-adrenoceptor (α1-AR) antagonists (Compounds B1 -B9) on α1-AR.METHODS ① A series of potential α1-adrenoceptor (α1-AR) antagonists,indolylpiperidine derivative (IPD) and Compounds B1 -B9,with indolylpiperidine moiety and different substitutes were synthesized through the coupling of indolylpiperidine and piperazine derivatives.② Inotropic responses experiment was used to examine blocking effects of IPD and Compounds B1 - B9 in isolated rat atria by phenylephrine (PE) stimulation.③ Blocking effect of IPD and Compounds B1 - B9 on phosphorylation level of extracellular signal-regulated kinase (ERK) in PE treated HEK293 cells was tested by Western blotting.RESULTS ① Potential α1-adrenoceptor (α1-AR) antagonists with indolylpiperidine moiety and different substitutes were synthesized successfully.② PE caused a dose-dependent inotropic response which was inhibited by pre-incubation of phentolamine (Phen),a non-selective α1-AR antagonist,IPD and Compounds B1,B3,B4,B7,B8 and B9,respectively; IPD and Compounds B4 and B8 caused an obvious rightward shift of inotropic response-curve,the pA2 values for IPD and Compounds B4 and B8 were 6.72 ± 0.21,6.86 ± 0.29 and 6.67 ± 0.19,respectively.③ Phosphorylation level of ERK1/2 was inhibited by pre-incubation with Compounds B1,B2,B3,B5,B6,B7,B8 and B9 or IPD in PE treated α1A-AR stably expressed HEK293 cells; PE-stimulated phosphorylation level of ERK1/2 was inhibited by pre-incubation with Compounds B2,B4,B7 or B8 in α1B-AR stably expressed HEK293 cells.CONCLUSION Compound B4 has a selective blocking activity on α1B-AR,and Compounds B1,B3,B5,B6 and B9 or IPD have a selective blocking activity on the phosphorylation level of ERK1/2.

Abstract: Objective To observe the phenotypic characteristics of 3 wild-type plague phages under different experimental environments, providing scientific evidence for the identification of phage biological characteristics and the study of their interaction with host bacteria in the future. Methods The sensitivity of 3 wild-type plague phages were detected by using liquid culture method, emisolid medium method and micro-liquid culture method based on OmniLog TM microbial identification system. Results The growth result based on LB liquid medium showed that the growth of plague phage 476 for 20-24 hours at both 28 ℃ and 37 ℃was better than that of plague phages 087 and 072204 at 37 ℃, and the growth of plague phages 087 was better than that of plague phages 072204 at 37 ℃. With the attenuated plague bacterium EV76 as the host bacterium, phage 476 was able to form visible plaque on double-layer agar medium for 20-20 hours at both 28 ℃ and 37 ℃, phages 087 and 072204 were only able to form opaque plaque on double-layer agar medium for 20-24 hours at 37 ℃. The growth results based on OmniLogTM system showed that when plague phage was lysed in EV76 strain at 33 ℃, the first row appeared as a straight line with a peak of no more than 100 in the 96-well microplate curve chart. As the phage quantity decreased, the dilution plate appeared with growth curve similar to EV76 strain in turn, and the color of tetrazolium dyes in the experimental wells gradually deepened as the phage number decreased and the host bacteria number increased. Therefore, it indicates that phage 476 was sensitively at both 28 ℃ and 37 ℃, while phage 087 and 072204 were temperature-dependent only at 37 ℃ to attenuated plague bacterium EV76. Conclusions The lysing ability of 3 wild-type plague phages are temperature-dependent, and the growth results are consistent under the three experimental conditions.

Objective To construct a technical platform for scarless gene modification of Yersinia pestis and to study the functions of its specific genes.Methods The resistance fragment, including upstream and downstream homologous arms of targeted regions, was reamplified by asymmetric PCR.The amplicons were introduced into Y.pestis harboring plasmid pKD46.With the induction of L-arabinose,the recombinant related enzymes: Exo, Beta and Gam, were expressed to guide the homologous recombination.A donor plasmid, pKSI-1, which carried the desired modification fragment flanking by I-SceⅠ recognition sites, was introduced into Y.pestis as the second step of λ-Red recombination with the help of pREDTKI.Results and Conclusion Two mutant strains:△waaA and waaA(△9nt), were successfully constructed for Y.pestis strain 201.Scarless modification introduces no extra modification to the genome, and it is ideal for comprehensive functional genomic studies.

Objective To explore the relationship between the size and location of the aneurysm after subarachnoid hemorrhage (aSAH) and its clinical classification. Methods A retrospective study was performed in patients with aSAH from January 1, 2008 to December 31, 2014. The relevant clinical data were collected including age, gender, aneurysm size, location, and Hunt-Hess (H-H) classification. The aneurysms were classified by size (A group d<5.00 mm, B group 5.00 mm≤d<10.00 mm, C group d≥10.00 mm), location and H-H classification according to the results of CT, digital subtrac?tion angiography (DSA), and magnetic resonance angiography (MRA). The relationship between size, position of aneurysm and H-H classification was observed and analyzed. Results There were 750 cases included in this study, with average age (56.14 ± 11.88), male 292 and female 458. The total number of aneurysms was 903, and the number of multiple aneurysms was 91 (12.13%). There was one case with multiple aneurysms that can be included in A, B and C groups. There were two cases with multiple aneurysms that can be included in A and B groups, two cases can be included in A and C groups, and three cases can be included in B and C groups. The number of aneurysms and the ratios of groups A, B and C were 20(3.9%), 12 (3.8%), 5 (7.5%), 70 (13.6%), 39 (12.2%), 10(14.9%), 2 (0.4%), 4 (1.3%), 2 (3.0%), 165 (32.0%), 94 (29.4%), 6 (9.0%), 130 (25.2%), 90 (28.1%), 6 (9.0%), 17 (3.3%), 11 (3.4%) and 2 (3.0%) for the location in the anterior cerebral artery, the middle cerebral artery, the posterior cerebral artery, the internal carotid artery, the anterior communicating artery, the posterior communicating artery, and the vertebral basilar artery, respectively. The number of aneurysms and the ratios of H-H classificationⅠ,Ⅱ,Ⅲ,ⅣandⅤin groups A, B and C were 48 (9.3%), 45 (14.1%), 12 (17.9%), 228 (44.2%), 150 (46.9%), 14 (20.9%), 68 (13.2%), 54 (16.9%), 30 (44.8%), 142 (27.5%), 43 (13.4%), 9 (13.4%), 30 (5.8%), 28 (8.8%) and 2 (3.0%). There was a negative correlation between the size of aneurysm and the H-H grade (rs=-0.075, P=0.024). Conclusion The anterior communicating artery and posterior communicating artery are high-risk areas for smaller aneurysms. The internal ca?rotid artery is high-risk areas for larger aneurysms. The size of aneurysm is negatively correlated with H-H classification.