2.In silico cloning and bioinformatics analysis of an AP2/EFR family gene from Arnebia euchroma.
Teng XIE ; Sheng WANG ; Liang-Yun ZHOU ; Jin-Fu TANG ; Lan-Ping GUO
China Journal of Chinese Materia Medica 2014;39(12):2251-2257
A cDNA sequence of Arnebia euchroma AP2/ERF named AeAP2/ERF was cloned by in silico cloning in this study, using ACX71873 sequence from Lithospermum erythrorhizon as the probe sequence. Some characters of the AP2/ERF gene and encoded protein sequences were predicted and analyzed by the bioinformatics methods, including general physical and chemical properties, hydrophobieity, signal peptide, secondary structure, localization sites in cells. Results showed that the 876 bp long gene included a 1 077 bp ORF and encoding 205 amino acid. The AeAP2/ERF protein had no signal peptide, it was a hydrophilic proteins located in nucleus. The function of the AP2/ERF protein was mainly involved with metabolism controlling and signal transduction.
Amino Acid Sequence
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Base Sequence
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Boraginaceae
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classification
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genetics
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Cloning, Molecular
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Computational Biology
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Computer Simulation
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Hydrophobic and Hydrophilic Interactions
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Molecular Sequence Data
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Phylogeny
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Plant Proteins
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genetics
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Protein Structure, Secondary
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Protein Structure, Tertiary
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Transcription Factors
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chemistry
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genetics
3.Quality evaluation of artificial musk based on its inhibitory effect on cyclooxygenase-2.
Yun LUO ; Cheng JIN ; Jian ZHOU ; Ruiqing WEN ; Xingfeng LI ; Ruisheng LI ; Ming YANG ; Xiaohe XIAO
Acta Pharmaceutica Sinica 2011;46(4):438-42
The inhibitory ratio (1%) of artificial musk on cyclooxygenase-2 (COX-2) was determined by enzyme immunoassay (EIA). The dose-effect relationship between concentrations of artificial musk and 1% was established. It was found that artificial musk had obvious inhibitory action on COX-2. The concentration for 50% of maximum inhibitory effect (IC50) was about 2.26 mg x mL(-1). There was a good relationship between the logarithm concentrations of artificial musk and 1% when the concentrations of artificial musk ranged from 0.31-20.0 mg x mL(-1). The results indicated that this EIA method could be applied to evaluate the anti-inflammatory activity of artificial musk quickly, conveniently, sensitively and exactly. This paper provided a novel method and foundational research for the bioassay of artificial musk.
4.The study of 921 and p53 expression in renal interstitial fibrosis
Yun XIAO ; Lijian TAO ; Tang DAMU ; Ou JIN ; Jianhua ZHOU ; Wangbing NING
Journal of Chinese Physician 2010;12(2):145-148
Objective To investigate the expression of p21 and p53 in renal interstitial fibrosis rats and the effect of enalapril on it. Methods Sprague-Dawley rats were randomly divided into 3 groups, shame operation rats group, unilateral urethral obstruction and enalapril treatment group. Histological chan-ges were observed by Masson stain. The expression of p21 mRNA and p53 mRNA was detected by RT-PCR. Results With degree of interstitial fibrosis aggravating, the expression of p21 and p53 increasing,p21 and p53 expression of UUO rats at every time point were positive correlative. Enalapril can inhibit the expression of p21 and p53. Concinsion p21 and p53 expression increased in UUO rats renal cortex and enalapril can significantly inhibit its expression, p21 may participate in the pathogenesis of renal tubule-in-terstitial fibrosis through p53 pathway.
5.The Expression of Plasma Membrane Ca2 + -ATPase Isoforms 1~4 and the Splice Variants at Sites A and C in the Neonatal Rat Vestibular Organ
Mi LUO ; Hanqi CHU ; Yanling TAO ; Liangqiang ZHOU ; Jin CHEN ; Yun LIU ; Chunchen PAN ; Qingguo CHEN
Journal of Audiology and Speech Pathology 2016;24(5):473-477
Objective To study the expression of plasma membrane Ca2 + -ATPase isoforms 1 -4 and the splice variants at sites A and C in the neonatal rat vestibular organ.Methods Ten rats at postnatal 2 days (P2 ) were decapitated and their vestibular organs (macula utriculi and macula sacculi)were isolated.The total proteins of the vestibular organs were extracted.The expression of PMCA1-4 splice variants at sites A and C was detected by RT-PCR.Results The splice variants of PMCA1-4 at sites A and C in macula utriculi and macula sacculi of neo-natal rat vestibular organs were PMCA1x/b,PMCA2w/(a,b),PMCA3z/(a,b,c)and PMCA4 (x,z)/b.Conclusion The splice variants at sites A and C among PMCA1,PMCA2,PMCA3 and PMCA4 were different in the vestibu-lar organs of neonatal rats,which could be explained that macula utriculi and macula sacculi had different require-ments of Ca2 + turning for these PMCA isoforms.
6.Expression of C2A domain of synaptotagmin Ⅰ fusion protein and its imaging in the ischemia-reperfusion rat model
Kang-yun, SUN ; Jun-dong, ZHOU ; Fei, SHEN ; Wei, FANG ; Jin-chang, WU
Chinese Journal of Nuclear Medicine 2010;30(5):312-315
Objective To evaluate myocardial apoptosis with 99Tcm-C2A-GST myocardial imaging using the recombined C2A domain of Synaptotagmin Ⅰ by gene engineering. Methods ( 1 ) The C2A gene was inserted into the prokaryotic glutathione S-transferate (GST) fusion protein expression plasmid pGEX-6P-1. The recombinant plasmid was transformed into E. coli BL21. C2A-GST fusion protein was purified after BL21 was induced with isopropyl-β-D-1-thiogalactopyranoside (IPTG). (2)The activity of fusion protein was identified by cell binding test with fluorescein-5-isothiocyanate (FITC)-C2A-GST. (3) The C2A-GST fusion protein was labeled with 99Tcm using 2-iminothiophene hydrocoride method. Radiochemical purity was determined with thin layer chromatography. (4)99Tcm-C2A-GST (7.4 MBq) was injected to ischemia-reperfusion rat models through tail vein. The image was acquired with SPECT at 1 h after injection, and then hearts were removed, rinsed with saline and dyed with triphenyl tetrazolium coride (TTC). The ischemic myocardium was separated from the viable myocardium and was weighted. Its radioactivity was measured by gamma counting. The difference of uptake of radiotracer between ischemic myocardium and normal myocardium was compared using percentage activity of injected dose per gram of tissue ( % ID/g) with standard deviation. SPSS 12.0 and t-test were used for data analysis. Results ( 1 ) C2A-GST fusion protein wassuccessfully expressed and its relative molecular weight was 3.8 × 104. (2) FITC-C2A-GST binding to apoptotic cells could be observed by fluorescent microscopy. (3) The radiochemical purity of 99Tcm-C2A-GST was (98.90 ±0.43)%. (4)The imaging studies showed that there was focal uptake of radioactivity in the ischemic myocardium. In vitro uptake of 99Tcm-C2A-GST was (2.41 ±0.32) % ID/g by the ischemic myocardium, however 99Tcm-C2A-GST-N-hydroxysuccinimide (C2A-GST-NHS) was (0. 82 ± 0. 24) % ID/g. There was statistically significant difference between those two groups (t = 10. 6, P <0.01 ). Conclusion The C2A domain of Synaptotagmin Ⅰ expressed by gene engineering can be used as the tracer for noninvasive detection of ischemic myocardium in the ischemia-reperfusion rat model.
7.Observation of Curative Effect ofHongteng Chang'an Decoction in Treating Ulcerative Colitis
Yun ZHOU ; Ji SUN ; Yaying WU ; Hongwei WANG ; Jing KONG ; Jin TANG
Chinese Journal of Information on Traditional Chinese Medicine 2015;(9):22-25
Objective To evaluate the clinical efficacy and safety ofHongteng Chang'an Decoction in treating mild and moderate ulcerative colitis (UC) cases.Methods Totally 120 patients of mild or moderate UC were randomly divided into two groups:TCM group of 60 patients, treated by self-madeHongteng Chang'an Decoction;Western medicine group of 60 patients, treated by mesalazine orally. The treatment course of both groups was 6 weeks, also with two-week follow-up. The ESR and CRP levels before and after treatment, the healing of colonic mucosa under endoscopy (according to the Baron score), the clinical efficacy and safety were analyzed statistically.Results CRP of both groups decreased significantly, with obvious statistical significance (P<0.05);ESR of both groups decreased, without statistical significance (P>0.05). The effects of healing of colonic mucosa under endoscopy of TCM group and Western medicine group were 61.37% (27/44) and 64.29% (27/42), respectively, without obvious statistical significance (P>0.05). The total effective rates of TCM group and Western medicine group were 86.7% (52/60) and 75.0% (45/60), respectively, and the TCM group was superior to the Western medicine group (P<0.05).ConclusionHongteng Chang'an Decoction can reduce the serum level of CRP of the mild and moderate UC patients effectively, relieve inflammation, and promote the healing of colonic mucosa. The treatment of mild and moderate ulcerative colitis byHongteng Chang'an Decoction has good clinical efficacy, without toxic side effects.
8.The Expression of Plasma Membrane Ca2 + -ATPase Isoforms 1 ~3in the Basi lar Membrane of Neonatal Rat Cochlea
Qingguo CHEN ; Hanqi CHU ; Liangqiang ZHOU ; Jin CHEN ; Yun LIU ; Mi LUO ; Yanling TAO
Journal of Audiology and Speech Pathology 2016;24(4):366-370
Objective To study the expression of plasma membrane Ca2+-ATPase isoforms 1~3 (PMCA 1~3 )in the basilar membrane (BM)of the neonatal rat cochlea by Western blot.The PMCA2 content in single BM of the neonatal rat was also examined.Methods Four rats at postnatal 2 days (P2)and 8 days (P8)were respective-ly decapitated and their BMs were isolated.The total proteins of BMs were extracted.The 20μg total proteins were respectively loaded to the gel.The expression of PMCA1-3 was detected by Western blot.Likewise,3μg total proteins from P2 and P8 rat BM were loaded.The expression of PMCA2 was detected by Western blot.Four rats at P8 were decapitated and their BM was isolated.The 5μg,10μg and 20μg total proteins of P8 rat BM were added to the gel and 100 ng,400 ng and 800 ng bovine serum albumin (BSA)were also loaded as reference.After electro-phoresis,the gel was separated into two parts.One part was used for SYPRO staining and the other part was used for PMCA2 detection by Western blot.Results In the 20μg BM total proteins of P2 and P8 rats,the expression of PMCA1 was weak (0.126±0.024,0.131±0.012,respectively),PMCA2 was strong (4.16±0.528,4.25±0.319, respectively),and PMCA3 was barely expressed (0 ).There was a statistical difference among PMCA1 ,PMCA2 and PMCA3(P<0.05).In the 3μg BM total proteins of P2 and P8 rats,the expression of PMCA2 in P8 (4.571± 0.336)was higher than P2 (3.622±0.285).There was a statistical difference(P<0.05).The PMCA2 content in the BM of a P8 rat was about 2 .5 ng.Conclusion There was a different-level expression of PMCA1~3 in the neonatal rat BM with highest expression of PMCA2 ,which could be explained that cochlear hair cells had different requirements of Ca2+ turning for these PMCA isoforms.
9.Study on the distinction of glycemic variability among different glucose regulation populations by phase space reconstruction of time series
Yun XIE ; Baoyi LI ; Xiaojun LUAN ; Jian ZHOU ; Liping HAN ; Jin LI ; Lele LIU ; Weiping JIA
Chinese Journal of Endocrinology and Metabolism 2012;28(9):722-725
Sixty-six volunteers,including 24 subjects with normal glucose tolerance(NGT),18 patients with impaired glucose regulation (IGR),and 24 patients with type 2 diabetes mellitus ( T2DM ),underwent a test of continuous glucose monitoring.The data of continuous glucose monitoring were embedded into two-dimension Euclid space by Takens' embedding theory.Glycemic phase diagram was drawn by MATLAB.The area and center distance of glycemic phase diagram were calculated by computer.The distinction of glycemic variability and average glycemic level among different glucose regulation populations were analyzed.The results showed that there existed significant differences in body mass index,systolic blood pressure,diastolic blood pressure,low density lipoprotein-cholesterol,high density lipoprotein-cholesterol,triglyceride,total cholesterol,creatinine,and alanine aminotransferase among three groups( all P<0.05 ).The levels of HbAIC,fasting plasma glucose( FPG ),postprandial 2 h plasma glucse (2hPG),area and center distance of glycemic phase diagram in T2DM group were higher than those in NGT and IGR groups( P<0.01 ),and the levels of FPG,2hPG,area and center distance of glycemic phase diagram in IGR group were higher than those in NGT group( P<0.01 ).The levels of FPG and 2hPG were correlated with area and center distance of glycemic phase diagram ( all P<0.01 ).These results suggest that measuring the area and center distance of glycemic phase diagram is a good method to assess glycemic variability and average glycemic level during continuous glucose monitoring.
10.Clinical analysis of congenital bile duct dilatation
Gang ZHOU ; Hong JIN ; Yun XIAO ; Tianrui ZHANG ; Yuping CHEN ; Zhiyong CHEN
Chinese Journal of General Surgery 2001;0(07):-
Objective To summarize the etiology,classification and clinical characteristics of congenital bile duct dilatation and our experience in its diagnosis and troatment.Methods The clinical data of 37 cases of congenital bile duct dilatation were retrospectively analyzed.Results The 37 cases included TypeⅠin 31 cases,Type Ⅱ in 1 case,Type Ⅳ in 4 cases,and Type Ⅴ in 1case.All of the patients underwent operative treatment.External drainage of was done in 2 cases,Roux-en-y cystojejunostomy in 3 cases,exeisison of cyst cyst and Roux-en-y hepaticojejunostomy in 31 cases and left hepatectomy in 1case with malignant change of cyst.Five cases had postoperative complications that recovered with non-operative treatment.Thirty-two cases were cured.There were no operative deaths.Conclusions Congenital bile duct dilatation should be treated as soon as possible.Excision of the cyst and Roux-en-y hepaticojejunostomy can reduce long-term postoperative complications and is the rational treatment for congenital bile duct dilatation.