Objective To express insulin-degrading enzyme(IDE)mutant T142A in prokaryotic cells and detect its activity.Methods According to the results of multi-sequence alignment and IDE substrate co-crystal structure,an active residue in β6-strand structure of IDE were predicted.The recombinant plasmid ppSUMO-T142A,with the site mutation of threonine 142 to alanine,was constructed by point mutation technique and expressed by E.coli prokaryotic expression system.After purification by nickel ion column affinity chromatography,ion exchange chromatography and gel filtration chromatography,the mutant T142A was obtained and determined for the activity by fluorescence method.Results IDE amino acid sequence is highly conserved among 16 species.T142 directly participates in substrate binding,interacts with substrate cleavage sites,and is close to important structures such as catalytic active sites and door-subdomains.The mutation of recombinant plasmid ppSUMO-T142A was proved to be correct by sequencing.The expressed fusion protein His-SUMO-T142A was mainly existed in soluble form in the supernatant at a concentration of 18 mg/mL,with a relative molecular mass of about 131 000;After three steps of purification,the purity of mutant T142A reached 86%.The maximum reaction rate(V_(max))of T142A catalytic degradation of fluorescent substrate V was 501.06 min~(-1) and the Michaelis constant(K_m) was 9.01μmol/L.Compared with wild-type IDE(V_(max) was 2 814.32 min~(-1),K_m was 11.93μmol/L),the activity of T142A decreased significantly.Conclusion The activity of IDE mutant T142A expressed in this study greatly decreases,while T142 is an important residue for IDE to play its enzymatic function,which provides an experimental basis for the development of new IDE activity regulatory molecules.

Cardiovascular Diseases ; Follistatin-Related Proteins ; physiology ; Humans

Objective: To study the chemical constituents of Artemisia anomala. Methods: Various chromatographic techniques were adopted to separate the constituents, and the spectroscopic analysis was used to identify their structures. Results: Seven compounds were isolated and identified as (4aS, 7S, 7aR)-methyl 7-hydroxy-7-methyl-1, 4a, 5, 6, 7, 7a-hexahydrocyclopenta [c] pyran-4- carboxylate (1), rehmaglutin D (2), (E)-6-hydroxy-2, 6-dimethylocta-2, 7-dienoic acid (3), chrysoeriol (4), luteolin (5), apigenin (6), and p-coumaric acid (7). Conclusion: Compound 1 is a new compound named artanoiridoid; Compound 2 is firstly reported in this genus; Compounds 3, 4, and 7 are separated from this plant for the first time.

The middle cerebral artery occlusion(MCAO) in rats remained a focus cerebral ischemic model that is non-incisioned,reliable,as well as the ischemic and refusion time can be controlled,which is widely used since it was created.However,because it is needed complex operations,associatied with many factors,it is difficult to establish.Many scholars have modified the operation,shape of the occlusion line,as well as the variety,narcotic drugs and so on.

China ; Clinical Laboratory Techniques ; Humans ; Myelodysplastic Syndromes ; diagnosis

Aortic Diseases ; Aortic Valve ; Aortic Valve Stenosis ; Calcinosis ; Humans ; Hyperlipoproteinemia Type II

Objective To establish a mathematical model to describe the skeletal class Ⅲ malocclusion of patient dental and basal bone arch form, for providing a data reference and basis for further study. Methods Thirty-five patients with skeletal classⅢmalocclusion were selected in this study for computed tomography CBCT. The data of 3-D image were analyzed, and dental arch marker (Fa) and base bone arch marker (Ba) were determined. The reference plane was determined by least square method. Software Matlab 7.0 was used to calculate two-dimensional coordinate system. Based on this, a mathematical model was established to describe the dental and basal bone arch form and then to validate the mathematical model. Results (1) The mathematical model can be used to describe the dental arch form of skeletal classⅢmalocclusion, maxillary:Y=46.12 [1-(2X/70.99)2]1.052;mandibular:Y=39.16 [1-(2X/64.51)2]1.038. (2) The mathematical model can be used to describe the basal bone arch form of skeletal classⅢmalocclusion, maxillary:Y=43.14 [1-(2X/75.09)2]1.061;mandibular:Y=39.03 [1-(2X/60.63)2]1.021. (3) Fa was located at Ba labial side in the maxilla, the distance was positive. Fa was located at Ba lingual side in the mandibular, and the distance was negative. (4) The fitting correlation coefficient of beta-function curve and each tooth on the dental and basal bone arch of skeletal class Ⅲ malocclusion were greater than 0.7 (P<0.05). Conclusion In this study, the mathematical model can be used to describe the dental and basal bone arch form of the skeletal classⅢmalocclusion, which can guide further research.

OBJECTIVE:To understand the application of proprietary Chinese medicine by clinicians in integrated medical insti-tutions,and provide reference for its rational use. METHODS:The questionnaire was designed,including related issues of proprie-tary Chinese medicine clinical application(such as medication basis of proprietary Chinese medicine,combined medication,clini-cal efficacy and its influencing factors,medication for special groups and so on)and suggestions;the questionnaires were delivered to the clinicians who worked in some second and third grade of integrated medical institutions located in southwestern regions and Chengdu,and the questionnaires was in the form of an anonymous;relevant data of available questionnaire were collected statisti-cally;the application situation of proprietary Chinese medicines and the existence question were analyzed,and suggestions were put forward. RESULTS:Totally 300 questionnaires were sent out,264 were effectively received,with effective rate of 88.0%. The clinicians involved in investigation were mainly in undergraduate,mostly the resident physicians and attending physician;only 6.8%of the physicians received the training of basic theory of traditional Chinese medicine after work;36.4%physicians often pre-scribe proprietary Chinese medicines,and the medication basis was mainly in package insert (64.0%) and clinical experience (55.3%);51.1% thought unclear indications affected the application of proprietary Chinese medicines,64.0% thought package in-sert can not meet clinical needs. In terms of answers in brief questions,the surveyed clinicians generally considered the package in-sert was too simple,which needed further perfected. CONCLUSIONS:The problems in clinical application of proprietary Chinese medicines should not be ignored. The relevant departments should strengthen the supervision of clinical application of proprietary Chinese medicines,and strengthen the re-evaluation of proprietary Chinese medicines market,as to provide data support for the continuous improvement of package insert. Medical institutions should carry out training timely for the physicians who prescribe pro-prietary Chinese medicines,the pharmacists should strengthen checking of proprietary Chinese medicines prescription and special prescription comment,and ensure prescription checking and comment play a active role in promoting rational drug use based on“proprietary Chinese medicines clinical application guiding principle”“prescription management method”.

Humans ; Leukemia, Large Granular Lymphocytic

Acute Disease ; Adolescent ; Adult ; Child ; Humans ; Leukemia ; diagnosis ; drug therapy ; Neoplasm, Residual ; Remission Induction