2.Expression and Preliminary Research on the Soluble Domain of EV-D68 3A Protein.
Ting LI ; Jia KONG ; Xiao-fang YU ; Xue HAN
Chinese Journal of Virology 2015;31(6):653-659
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
4.Correlation between complete denture relines and serum bone turnover markers in elderly edentulous patients with osteoporosis
Tianhao YU ; Xingya JIA ; Yanqing XUE ; Desong ZHAN ; Xu YAN
Chinese Journal of Tissue Engineering Research 2017;21(22):3513-3518
BACKGROUND:The residual alveolar ridge in edentulous patients with osteoporosis always presents with a poor shape,which increases the difficulty in complete denture restoration.Until now,there are no predictors for residual alveolar ridge resorption or guidelines on the frequency of denture relines.OBJECTIVE:To investigate the correlation between the frequency of complete denture relines and serum bone turnover markers,osteocalcin (OC) and cross-linked carboxy-terminal telopeptide of type Ⅰ collagen (CTX),in elderly edentulous patients with osteoporosis,in order to provide reference for complete denture repair.METHODS:According to inclusion criteria,50 elderly edentulous patients with osteoporosis who had complete dentures were recruited in the study randomly.The frequency of complete denture relines was surveyed by related questionnaire and medical record,and the serum OC and CTX levels were detected by electrochemiluminescence immunoassay.Related data were analyzed statistically via Pearson correlation analysis.RESULTS AND CONCLUSION:It showed a strong positive correlation between the frequency of complete denture relines and the serum OC level (r=0.517,P < 0.01).No significant correlation between relines frequency and the serum CTX level was observed (r=0.278,P=0.051 > 0.05),but it showed a similar tendency between them.These findings indicate that to detect the levels of serum bone turnover markers may be conducive to evaluating curative effect of complete dentures in elderly edentulous patients with osteoporosis as well as to making subsequent diagnostic and therapeutic strategies.
5.Epidemiological features of infectious disease related public health emergencies in Zhejiang Province, 2010-2018
ZHAI Yu Jia ; LIN Jun Fen ; GU Xue ; HE Fan
Journal of Preventive Medicine 2021;33(1):21-24
Objective:
To learn the epidemiological characteristics of infectious disease related public health emergencies in Zhejiang Province from 2010 to 2018 for the prevention and control.
Methods:
The surveillance data was extracted from National Public Health Emergency Management Information System. Descriptive epidemiology method was used to analyze main diseases as well as distribution characteristics of time and places.
Results:
A total of 445 events were reported, which caused 14362 cases and 34 deaths, with a attack rate of 0.69% and mortality rate of 0.24%. There were 298 events with less than 30 cases, accounting for 66.97%. The event classification was dominated by general events ( 242 events, 54.38% ) and ungraded events ( 201 events, 45.17% ). The main diseases were chickenpox ( 134 events, 30.11% ), hand foot mouth disease ( 59 events, 13.26% ) and other infectious diarrhea ( 51 events, 11.46% ). The incidence peaked from April to June ( 129 events, 28.99% ) and from November to December ( 131 events, 29.44% ). Ningbo ranked the top in the number of reported events ( 141 events, 31.69% ). Most events ( 322 events, 72.36% ) occurred in schools.
Conclusions
The infectious disease related public health emergencies in Zhejiang Province from 2010 to 2018 were mainly caused by chickenpox, hand-foot-mouth disease and other infectious diarrhea. The two peaks of the emergencies occurred from April to June and from November to December. Ningbo was the main area reporting infectious diseases, and schools were the main places.
6.The influence of rhAng-1 on endothelial junctions of BBB after focal cerebral ischemia/reperfusion in rats.
Hang YU ; Qing-Bo JIA ; Yi-Xue XUE
Chinese Journal of Applied Physiology 2013;29(2):147-152
OBJECTIVEThis study was performed to determine whether recombinant human angiopoietin-1 (rhAng-1) decreases the permeability of the blood-brain barrier (BBB) in focal cerebral ischemia and reperfusion rats, whether RhAng-1 opens the BBB by affecting tight junction associated proteins zonnula occludin-1 (ZO-1), occludin and adherens junction protein vascular endothelial (VE)-cadherin.
METHODSThe rats were divided into eight groups randomly( n = 10): (1) sham-operated group; (2) ischemia group; (3)-(5) ischemia/reperfusion (middle cerebral artery occlusion and reperfusion (MCAO/R) 12 h, 48 h, and 7 days) and 0.9% saline groups; (6)-(8) ischemia/reperfusion (MCAO/R 12 h, 48 h, and 7 days) and rhAng-1 groups. The Bee permeability was assessed by Evans blue extravasation. The messenger RNA and protein expressions of ZO-1, occludin, and VE-cadherin were determined by reverse transcription-polymerase chain reaction, Western blot, and immunohistochemistry assays.
RESULTSThe BBB permeability and the brain infarct volume were significantly decreased after rhAng-1 injection. The expressions of ZO-1, occludin, and VE-cadherin were increased after rhAng-1 injection.
CONCLUSIONrhAng-1 may decrease the permeability of BBB in MCAO/R rats by upregulation of ZO-1, occludin, and VE-cadherin.
Angiopoietin-1 ; pharmacology ; Animals ; Antigens, CD ; metabolism ; Blood-Brain Barrier ; cytology ; metabolism ; Brain Ischemia ; metabolism ; Cadherins ; metabolism ; Endothelium, Vascular ; cytology ; drug effects ; Humans ; Male ; Occludin ; metabolism ; Rats ; Rats, Wistar ; Recombinant Proteins ; pharmacology ; Reperfusion Injury ; metabolism ; Zonula Occludens-1 Protein ; metabolism
7.Development of core competence evaluation system for preventive medicine undergraduates based on Delphi method
Xuanyue TANG ; Xiaoqiu LIU ; Jia YU ; Xue LIU ; Litian SHI ; Li JIA
Chinese Journal of Medical Education Research 2017;16(1):17-22
Objective To develop a core competence evaluation index system for preventive medicine undergraduates.Methods Following the scientific,systematic and applicable principles,we proposed evaluation index system of core competence for professional medical prevention and selected 35 experts for three rounds expert consultation on the index system by using Delphi method.The basic situation,the positive and authority degree of experts and all levels of index weight coefficient were analyzed using SPSS 13.0 software.Results The core competence evaluation index system for preventive medicine undergraduates included six dimensionalities,fifteen secondary indicators and forty-nine tertiary indicators.The six dimensionalities included professional value behavior (0.25),professional knowledge and skills (0.28),basic specialized knowledge (0.20),communication (0.13),information management (0.06) and systems thinking (0.08).Conclusion The establishment of the evaluation system can provide a scientific and objective reference for the evaluation of the core competence of medical students.
8.Study on the Quality Standard of Tongmai Pill
Sijiao CUI ; Shilong YU ; Qiang XUE ; Guangzhao YANG ; Wenyu ZHONG ; Jingyang XU ; Jia NIU ; Yu WANG
China Pharmacy 2016;(6):821-823
OBJECTIVE:To establish the quality standard for Tongmai pill. METHODS:TLC was used for the qualitative iden-tification of Salvia miltiorrhiza,Angelica sinensis and Carthamus tinctorius. HPLC was used for the content determination of feru-lic acid. The column was Diamonsil C18 with mobile phase of acetonitrile-0.1% aqueous phosphoric acid (17∶83,V/V) at a flow rate of 1 ml/min,the detection wavelength was 316 nm,the column temperature was 30 ℃,and the injection volume was 10 μl.. RESULTS:The TLC of S. miltiorrhiza,A. sinensis and C. tinctorius showed clear spots and good separation. The linear range of ferulic acid was 8-80 μg/ml(r=0.999 8);RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 97.2%-99.6%(RSD=1.2%,n=6). CONCLUSIONS:The standard can be used for the quality control of Tongmai pill.
9.Effects of Iron Chelators on Labile lron Pool and Apoptosis Related Genes Ex pression in K562 Cells
xue-qiang, WU ; guo-cun, JIA ; yi-ming, YANG ; yu-feng, LIU ; yu-ren, XI
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To explore the effect of iron chelators on labile iron pool and expression of apoptosis associated genes in cells of K562, an erythroleukemia cell line.Methods K562 cells were incubated at 37 ℃ in RPMI 1640 containing 10% heat-inactived fetal bovine serum in an saturated humidity and 5% CO_2 incubator. K562 cells were incubated with different concentrations of desferro-(xamine(DFO)). The study groups were divided as following: DFO group, iron+DFO group and the control group. Following indices were detected which included apoptosis by flow cytometry (FCM) assay, expression of Rb, c-myc, bax mRNA by RT-PCR. The intracellular LIP was measured with a fluorimetric assay using the metalsensitive probe calcein-AM.Results 1. The viability of K562 cells incubated with different concentrations of DFO was lower than that of control group at 12 h,24 h and 48 h (P
10.Controlled observation on the efficacy of thoracic facet joint disorder treated with electroacupuncture and manual reduction.
Tian YE ; Hong-Wei XUE ; Yu WANG ; Lan LIU ; Jia-Bo SUN
Chinese Acupuncture & Moxibustion 2013;33(12):1077-1080
OBJECTIVETo compare the difference in the efficacy on thoracic facet joint disorder between the combined therapy of electroacupuncture and manual reduction and the simple manual reduction.
METHODSOne hundred and sixty patients were randomized into an electroacupuncture and manual manipulation group (group A) and a simple manual manipulation group (group B), 80 cases in each one. In the group A, Ashi points and three pairs of Jiaji (EX-B 2) bilateral to the painful sites were selected. The perpendicular puncture was used at Ashi points, the oblique puncture was used at Jiaji (EX-B 2) and connected with electric stimulation for 20 min, additionally, the corresponding manual reduction was adopted at the sites of facet joint disorder. In the group B, the simple manual reduction was applied to the affected sites. Acupuncture was given once every day, the manual reduction was applied once every 10 days. The treatment of 10 days made one session. The efficacy was analyzed statistically at the end of two sessions of treatment. Before and after treatment, McGill pain scale was adopted for the value statistical analysis. PRI score, VAS score and PPI score of patients were calculated before and after treatment and compared in the two groups. The efficacy was compared between the two groups.
RESULTSThe curative rate was 56.3% (45/80) in the group A, which was better than 18.8% (15/80) in the group B (P< 0.01). The total effective rate was 95.0% (76/80) in the group A, which was better than 76.3% (61/80) in the group B (P<0.01). The scores of PRI, VAS and PPI after treatment were all improved significantly in the two groups (all P<0.05), in which, the results in the group A were better than those in the group B (PRI: 4.00 +/- 0.97 vs 5.44 +/- 1.16, VAS: 3.29 +/- 0.72 vs 3.87 +/- 0.81, PPI: 1.07 +/- 0.74 vs 1.64 +/- 0.90, all P<0.05).
CONCLUSIONThe combined therapy of electroacupuncture and manual manipulation achieves the superior efficacy on thoracic facet joint disorder as compared with the simple manual manipulation. The combined therapy relieves the symptoms of thoracic facet joint disorder and reduces the severity of disorder.
Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Thoracic Diseases ; therapy