To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism

Asthma is a common disease with recurrent onset which severely affects patients' quality of life.Acupuncture can improve pulmonary functions in asthma patients and thus treat this disorder.To summarize the status of acupuncture treatment for asthma,we have collected clinical literatures published in the recent 10 years and analyzed the influence of acupuncture on pulmonary functions in asthma patients from the aspects of frequently used points,needling techniques,manipulation and mechanisms to provide references for treating asthma with acupuncture.

Adult ; Female ; Humans ; Magnetic Resonance Imaging ; Neuroma, Acoustic ; pathology ; surgery

AIMTo study the chemical constituents of Lagopsis supina.

METHODSCompounds were isolated by column chromatography of silica gel and Sephadex LH-20, and the structures were determined by spectral analysis.

RESULTSTwo compounds were isolated and elucidated as apigenin-7-O-(6"-(E)-p-coumaroyl)-beta-D-galactopyranoside (I) and apigenin-7-O-(3",6"-di-(E)-p-coumaroyl)-beta-D-galactopyranoside (II).

CONCLUSIONI and II are new compounds.

Apigenin ; Flavonoids ; chemistry ; isolation & purification ; Galactosides ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Molecular Structure ; Plants, Medicinal ; chemistry

?AlM: To investigate the expression of miR-181 in the lens tissue of cataract and the regulating mechanism of miR-181 on apoptosis of human lens epithelial cell.
?METHODS:Real time q-PCR was used to measure the expression of miR-181 in the anterior lens capsules of age - related cataract and human lens epithelial cell apoptosis model. miR- 181 mimic and inhibitor were transfected using Lipofectamine 2 000 to regulate the expression of miR-181, and then Real time q-PCR was used to verify transfection efficiency. Flow cytometry was used to detect the change of cell apoptosis rate.
? RESULTS: Compared with control group, the expression of miR-181 was significantly higher in both the anterior lens capsules of age-related cataract and human lens epithelial cell apoptosis model; the relative expression of miR-181 in lens epithelial cells transfected with miR-181 mimic was increased, whereas decreased in cells transfected with miR-181 inhibitor;the apoptosis rate of cells transfected with miR - 181 mimic was increased, while reduced in miR-181 inhibitor group. Each result was statistically significant (P<0. 01).
?CONCLUSlON:High expression of miR-181 is detected in anterior lens capsule of age-related cataract. miR-181 might play a certain role in the pathogenesis of cataract via promoting human lens epithelial cell apoptosis. miR-181 probably becomes a new approach for the nonoperative treatment of cataract, but the concrete mechanism still needs to be further studied.

Objective To evaluate the performance of the ABX Micro C-reactive protein(CRP)in determination of CRP.Methods The analytic characteristics including precision,carry-over,linearity, stability,interference and comparability were examined.Results The coefficient of variation(CV)was less than 5.1%,10% and d.3% for within-run,between-run and between-day,respectively.Carryover was less than 1.2%.Whole blood samples held at either room temperature or 4℃ were stable for 48 hours with relative deviation less than 6.0% relatively.Linear range was 1.0-70.0 mg/L using undiluted samples.The comparison between the ABX Micro CRP and Behring Nephelometer Ⅱ was well correlated Both serum:Y=0.996 7X-0.398 5,r~2=0.965 9;serum for BN Ⅱ,whole-blood samples for the ABX Micro CRP:Y=0.908 8X-0.138 2,r~2=0.959 4;both serum and whole-blood samples for the ABX Micro CRP: Y=1.001 7X-0.898 2,r~2=0.952 7.No obvious interference was observed by hyperhemoglobinemia and hyperlipidemia.Conclusion The determination of CRP test with ABX Micro is accurate and reliable.

Harmful substances are produced during the metabolism of alcohol in the human body, which causes liver disease.The mechanism of oxidative stress is the main pathogenesis of alcoholic liver disease, which plays a major role in the occurrence and development of alcoholic liver disease.Alcohol-induced oxidative stress mediated by reactive oxygen species(ROS)/reactive nitrogen species(RNS) brings about liver disease via apoptotic signaling pathway and MAPK signaling pathway, and alleviates liver disease through Nrf2 signaling pathway.In this paper, the studies on the mechanism of oxidative stress in alcoholic liver disease are reviewed.

Objective:To understand the patients'satisfaction on catastrophic medical insurance of New Rural Cooperative Medical Scheme ( NRCMS ) , and provide references for effective management and policy improvement. Methods:According to the principles of representative sampling, a questionnaire survey conducted. 484 patients got compensation for catastrophic insurance in 2013 were surveyed, and information of medical costs was analyzed. Re-sults:The average medical cost accounts for 84. 57% of total household expenditure. Mandatory medical expenditure is still high and economic burden of diseases is heavy. This policy has increased the compensation rate, and the actu-al compensation rate increases 4. 79% in three districts. The average score of patients' attitude toward catastrophic medical insurance is (3.95±1.05) points, 88. 43% of patients continue to participate in medical insurance. Con-clusions and suggestions:Patients' satisfaction on catastrophic medical insurance of NRCMS is high. It is suggested that the existing financing modes should maintain stability in the short term, and the compensatory rate should be im-proved reasonably.

This study aimed to identify the related substances of phloroglucinol injection by two-dimensional liquid chromatography quadrupole time-of-flight mass spectrometry (2D-LC-Q-TOF/MS). The first-dimensional separation was carried out on an HSS T3 (250 mm × 4.6 mm, 5 μm) column by gradient elution using 1.36 g·L^-1 potassium dihydrogen phosphate buffer solution (pH adjusted to 3.0 with diluted phosphoric acid) and acetonitrile as the mobile phases. The separated components were then trapped in switch valve tube lines respectively and delivered to the second-dimensional desalting gradient elution which was performed with a BDS C18 (100 mm × 4.6 mm, 2.4 μm) column using 0.1% formic acid and methanol as the mobile phases. After rapid desalting, electrospray-ionization quadrupole time-of-flight high resolution mass spectrometry was used for determining the accurate masses and elemental compositions of the parents and their product ions for both phloroglucinol and its related substance. Structures of the related substances were then figured out by mass spectrometry elucidation, organic reaction mechanism analysis, and/or comparison with reference substances. Under the established analytical conditions, phloroglucinol and its related substances were adequately separated, 17 main related substances were detected and identified in the injection and its stressed samples for the first time. The identification results can provide reference for the quality control of phloroglucinol injection.

To study the chemical constituents from the the deep-sea fungus Alternaria sp. F49. Seven compounds were isolated from the EtOAc extract by using silica gel, Sephadex LH-20, ODS and HPLC methods. Based on the spectroscopic analysis, their structures were identified as (8R)-5-O-methyl-orcinotriol (1), orcinotriol (2), α-acetylorcinol (3), 3'-hydroxyalternariol 5-O-methyl ether (4), altenusiol (5), altenusin (6), and 5'-methoxy-6-methyl-biphenyl-3,4,3'-triol (7). (8R)-5-O-Methyl-orcinotriol (1) is a new phenolic compound which has never been reported in the literature. Compounds 4-7 showed strong DPPH free radical scavenging activity; whereas compounds 1-7 showed strong ABTS free radical scavenging activity.