1.Crystal structures of plant uridine diphosphate-dependent glycosyltransferases.
Heshu LÜ ; Feiyan XUE ; Chunmei LIU ; Mingfeng YANG ; Lanqing MA
Chinese Journal of Biotechnology 2014;30(6):838-847
Glycosyltransferases (GTs) catalyze the transfer of a sugar residue of an activated sugar donor to an acceptor molecule. Many families 1 GTs utilize an uridine diphosphate (UDP) activated sugar as donor in the glycosylation reaction, and most of these belong to a group of GTs referred to as the UGTs. The relationship between the degree of amino acid sequence identity and substrate specificity of the plant UGTs is highly complicated, and the prediction of substrate specificity based on phylogenetic analyses need to be improved by more biochemical characterization. This review summarizes the three dimensional structures of plant UGTs published in the Protein Data Bank (PDB), including the detailed substrate interactions with the sugar and receptor binding pockets and mutational analyses of some critical amino acids. It will be helpful for biochemical characterization the substrate specificity of the individual UGT, and lay the foundation for the enzymatic and genetic manipulation of plant UGTs in the future.
Amino Acid Sequence
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Glycosylation
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Glycosyltransferases
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chemistry
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Phylogeny
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Plant Proteins
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chemistry
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Plants
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enzymology
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Protein Structure, Tertiary
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Substrate Specificity
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Uridine Diphosphate
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chemistry
2.Effect of Human Hepatocellular Carcinoma HepG2 Cell-derived Exosome on the Differentiation of Mesenchymal Stem Cells and Their Interaction.
Fei LUO ; Zhao SUN ; Qin HAN ; Chunling XUE ; Chunmei BAI
Acta Academiae Medicinae Sinicae 2017;39(3):312-317
Objective To investigate the effect of human hepatocellular carcinoma HepG2 cell-derived Exosome on the differentiation of mesenchymal stem cells(MSC)into cancer-associated myofibroblasts(CAF)and the impacts of CAF on liver cancer cell proliferation,migration,and invasion. Methods The protein expression of HepG2 cell-derived Exosome was detected by Western blotting. MSCs were separated from human adipose tissue and cultured with HepG2 cell-derived Exosome(100 ng/nl)to initiate differentiation. The expressions of mesenchymal markers and several interleukins were also detected by Western blotting. HepG2 cells were co-cultured with the conditioned media(CM),in which HepG2 Exosome induced the differentiation of MSC into CAF. The expressions of epithelial and mesenchymal markers were detected by real-time polymerase chain reaction(PCR)and Western blotting. Cell proliferation was assessed using MTS assay. Transwell chambers were used in the in vitro migration and invasion assay. Results HepG2 cell-derived particles expressed CD63,70 kilodalton heat shock proteins,and 90 kilodalton heat shock proteins. With the treatment of HepG2 cell-derived Exosome,the expressions of mesenchymal marker α-smooth muscle actin,fibroblast activation protein α,interleukin(IL)-6,IL-8,and IL-1β were up-regulated,while vascular endothelial growth factor had no significant change. The conditioned media which HepG2 Exosome induced MSC differentiation CAF(CAF-CM)could significantly promote HepG2 cells proliferation(1.075±0.104),compared to BSA control(0.874±0.066,P=0.023)and MSC-CM(0.649±0.034,P=0.0005). CAF-CM could significantly enhance cell migration [(42.5±9.1) cells vs.(18.5±3.1) cells,P=0.001] and invasion [(29.0±3.5) cells vs.(13.1±3.7) cells,P=0.009] compared to its control group. Moreover the conditioned medium which HepG2 Exosome induced MSC to differentiate into CAF could also promote the expressions of mesenchyme-related genes Smad interacting protein 1(P=0.040),β-catenin(P=0.038),fibronectin(P=0.029),and Vimentin(P=0.013)and inhibit the expression of epithelial related genes zonula ocdudens-1(P=0.010).Conclusions Exosome extracted from HepG2 cells can induce human adipose-derived MSC to differentiate into cancer-associated myofibroblasts. CAF-like cells can promote the migration of the liver cancer cell line HepG2.
3.The correlation between matrix metalloproteinase-9 and cardiac function,aneurysm volume in the process of ventricular aneurysm formation
Hong ZHAI ; Chunmei WANG ; Lina GUAN ; Xue YAN
Chongqing Medicine 2015;(32):4487-4489
Objective To explore the correlation between matrix metalloproteinase‐9(MMP‐9) and cardiac function ,aneu‐rysm volume in the process of left ventricular aneurysm (LVA) formation .Methods Rabbit models of LVA were established in 20 New Zealand rabbits by lighting left anterior descending artery and left circumflex artery .Two‐dimensional echocardiography were performed at preoperative and postoperative 1 d ,2 d ,3 d ,1 w ,2 w ,3 w ,4 w .The survived animals which had LVA formed were en‐rolled .Real time three‐dimensional echocardiography was performed to obtain left ventricular volume ,LVEF and LVA volume ,The MMP‐9 serum concentration was measured by ELISA .Results (1)There were a significant increase in the serum concentration of MMP‐9 from postoperative 1 d and arrived at the peak at postoperative 3 d ,there were higher than preoperative at postoperative 4 w (P<0 .05) .(2)Compared with preoperative ,the LVEDV ,LVESV and LVAV volume at postoperative showed an increase trend (P<0 .05) ,while there was a decreasing trend in LVEF(P<0 .05) .(3)The MMP‐9 had the better relationship with LVEF at post‐operative 1 d ,2 d ,3 d ,1 w ,2 w ,3 w ,4 w ,in which MMP‐9 had the tightest relationship with LVEF at postoperative 3 d (r=0 .731 , P<0 .05) .MMP‐9 had the better relationship with LVA volume at postoperative 2 d ,3 d ,1 w ,2 w ,3 w ,4 w ,in which MMP‐9 had the tightest relationship with LVA volume at postoperative 3 d (r=0 .636 ,P<0 .05) .Conclusion The MMP‐9 had an effect on cardiac function and LVA volume in the process of LVA formation .
4.Clinical Observation of Intranasal Dexmedetomidine and Sufentanil for Pediatric Sedation Undergoing Stomatology Operation of Outpatient Department
Jing CHU ; Hong LI ; Chunmei CAI ; Peitong XUE ; Jinrong HUO
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(3):438-442
[Objective] To research the sedative effect of intranasal dexmedetomidine and sufentanil for pediatric sedation for stomatological operation of outpatient department.[Methods] 60 children undergoing stomatological operation of outpatient department,age 3 ~ 7 years,weighing 10 ~ 32 kg,of ASA physical status Ⅰ ~ Ⅱ,were divided into the three groups (n =20) randomly using a random number table:group dexmedetomidine and sufentanil (group DS),group dexmetomidine (group D),group sufentanil (group S).Recorded the children's behavior using the Ohio State University behavior rating score (OSUBRS),the University of Michigan Sedation Score (UMSS),SBP,HR and side-effects when entry,during and leave operation and in post-anesthesia care unit,side-effects,the satisfaction of stomatological doctors and parents.[Results] The OSUBRS of group DS when entry,during operation were lower than group D (P < 0.01).The UMSS of group DS were higher than group D and group S when entry and during operation (P < 0.05).The success rate of group DS was higher than group D and group S (P < 0.01).There was no different of mean percentage change in systolic blood pressure and heart rate from baseline between group DS and group D (P > 0.05).There were no instances of respiratory depression,hypotension and bradycadia.[Conclusion] Intranasal dexmedetomidine and sufentanil provides satisfactory pediatric sedation for stomatological operation of outpatient department without side effects such as respiratory depression,bradycadia and hypotension.
5.Influence of nursing health education on morning blood pressure surge fluctuations in aged people with hypertension
Qian WEN ; Xue BAI ; Lingling HUANG ; Chunmei XU
Chinese Journal of cardiovascular Rehabilitation Medicine 2017;26(2):210-213
Objective: To explore influence of nursing health education on morning blood pressure surge (MBPS) fluctuations in aged patients with hypertension.Methods: A total of 100 outpatients and inpatients with non-dipper hypertension, who haven't been treated, were selected from Apr 2011 to Sep 2015.According to random number table method, patients were randomly and equally divided into routine treatment group and health education group (received nursing health education based on routine treatment group), both groups were treated for four weeks.MBPS, trough/peak(T/P) ratio and smoothness index (SI) were measured and compared between two groups after treatment.Results: Compared with routine treatment group after four-week treatment, there were significant reductions in systolic blood pressure (SBP) morning surge[(46.95±5.17) mmHg vs.(42.21±5.19) mmHg]and diastolic blood pressure (DBP) morning surge[(16.31±3.23) mmHg vs.(14.21±3.58) mmHg], and significant rise in SBP T/P[(59.22±6.65)% vs.(70.11±6.41)%], DBP T/P[(50.91±5.04)% vs.(56.97±5.12)%], SBP SI[(1.19±0.13) vs.(1.42±0.12)]and DBP SI[(1.11±0.09) vs.(1.29±0.08)]in health education group, P<0.05 all.Conclusion: Health education can effectively improve blood pressure fluctuations and may likely improve prognosis in patients with non-dipper hypertension, which is worth extending.
6.Terminal Bacterial Pollution of Hospital Central Oxygen Supply Equipment and Sanitizing Effect of MX-1 Disinfector
Xin XUE ; Zhenshan WANG ; Xiaohong AN ; Chunmei YANG ; Junming GU
Chinese Journal of Nosocomiology 2006;0(03):-
OBJECTIVE To investigate the terminal bacterial pollution of hospital central oxygen supply system and the sanitizing effect of MX-1 disinfector. METHODS Timing many spot samples of terminal oxygen supply equipment (oxygen flowmeter, humid bottle, oxygen pipe) were quantified and cultivated through 24 hours continue supply. RESULTS Five cases of timing many spot samples testified that there were no bacteria oxygen pollution in the pipe oxygen supply in the Department of Respiratory Disease, the Second Affiliated Hospital of Dalian Medical University, and that there were no bacteria pollution in the terminal oxygen supply equipments if they were sterilized strictly, but there would be severe bacteria pollution if they were not sterilized strictly. This test found that lid of humid bottle and lapis were the main source of bacteria pollution. Connection with MX-1 disinfector in the terminal can avoid bacteria pollution. CONCLUSIONS Severe bacteria pollution will happen if the oxygen supply equipments aren't sterilized strictly; connection with MX-1 disinfector in the terminal can avoid bacteria pollution and ensure oxygen for cleanliness and safety.
7.Preparation and crystallization of Polygonum cuspidatum benzalacetone synthase.
Wenrui MA ; Chunmei LIU ; Mingfeng YANG ; Feiyan XUE ; Qing CHEN ; Lanqing MA ; Heshu LÜ
Chinese Journal of Biotechnology 2016;32(2):250-258
The chalcone synthase (CHS) superfamily of the type III polyketide synthases (PKSs) generates backbones of a variety of plant secondary metabolites. Benzalacetone synthase (BAS) catalyzes a condensation reaction of decarboxylation between the substrates of 4-coumaric coenzyme A and malonyl coenzyme A to generate benzylidene acetone, whose derivatives are series of compounds with various biological activities. A BAS gene Pcpks2 and a bifunctional CHS/BAS PcPKSI were isolated from medicinal plant P. cuspidatum. Crystallographic and structure-based mutagenesis studies indicate that the functional diversity of the CHS-superfamily enzymes is principally derived from small modifications of the active site architecture. In order to obtain an understanding of the biosynthesis of polyketides in P. cuspidatum, which has been poorly described, as well as of its activation mechanism, PcPKS2 was overexpressed in Escherichia coli as a C-terminally poly-His-tagged fusion protein, purified to homogeneity and crystallized, which is helpful for the clarification of the catalytic mechanism of the enzyme and lays the foundation for its genetic engineering manipulation.
Butanones
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Catalytic Domain
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Crystallization
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Fallopia japonica
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enzymology
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Polyketide Synthases
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genetics
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metabolism
8.Association of OSAS in patients with type 2 diabetes with levels of tumor necrosis factor-α and plasminogen activator inhibitor
Hongxia ZHU ; Zhenshan WANG ; Xin XUE ; Ping ZHANG ; Chunmei YANG ; Benli SU
Chinese Journal of Endocrinology and Metabolism 2013;29(8):679-682
Objective To investigate the changes in tumor necrosis factor-α(TNF-α) and plasminogen activator inhibitor(PAI) in the presence of obstructive sleep apnea syndrome(OSAS) in patients with type 2 diabetes mellitus,and to investigate if that 2 inflammatory factors is associated with hypoxemia during sleep.Methods Fiftyfour patients with type 2 diabetes mellitus hospitalized for glycemic control were subgrouped into 27 patients with OSAS (group O) and 27 patients without OSAS (group N).26 people without diabetes and OSAS were selected from a health check-up program to serve as control group (group C).Serum TNF-α and PAI levels were measured by commercial ELISA kits.Apnea hypopnea index (AHI),the lowest pulse oxygen saturation (LSpO2) at night were detected by portable sleep monitor.The relativity regarding TNF-α,PAI,homeostasis model assessment for insulin resistance (HOMA-IR),fasting blood glucose(FBG),AHI,LSpO2,and age was discussed.TNF-α and PAI were regard as dependent variables,and FBG,HOMA-IR,AHI,LSpO2,and age as independent variables in multivariate stepwise linear analysis.Results The levels of HOMA-IR,TNF-α,PAI in group O [2.69 ± 1.53,(1.97 ±0.13) ng/ml,and (2.27 ± 0.85) ng/ml] were higher than the those in other 2 groups,and those in group N [1.70 ± 0.88,(1.09 ±0.29) ng/ml,and (1.59 ± 0.13) ng/ml] were higher than group C [1.15 ± 0.67,(0.73 ± 0.19) ng/ml,and (0.89 ±0.25) ng/ml].The levels of FBG in the 2 diabetic groups were higher than those in group C.The level of AHI in group O was higher than that in group N and group C,while LSpO2 in group O was lower than that in group N and group C.Spearman univariate analysis showed that TNF-α and PAl both were associated positively with HOMA-IR,FBG,and AHI,while negatively with LSpO2.Multivariate stepwise linear analysis denoted that TNF-α was independently associated with AHI,FBG,while PAI was independently associated with AHI and HOMA-IR.Conclusions Patients with diabetes and OSAS manifest raised level of chronic inflammatory activity,and therefore,OSAS might be implicated in the pathogenesis of chronic complication of patients with type 2 diabetes.
9.Expression of miR-155 in peripheral blood and skin lesions from as well as its relationship with Th17 cells in patients with atopic dermatitis
Lei MA ; Haibo XUE ; Xiuhao GUAN ; Chunmei SHU ; Juan YU ; Junhua ZHANG ; Beilei YUN
Chinese Journal of Dermatology 2014;47(1):15-18
Objective To detect the expressions of miR-155,T helper type 17 (Thl7) cells,and Th17 cellspecific transcription factor RORγt and effector cytokine interleukin (IL)-17 in peripheral blood and skin lesions from,and to evaluate their relationship in,patients with atopic dermatitis (AD).Methods Peripheral blood was obtained from 37 patients with AD and 33 age-and sex-matched healthy controls,and biopsy specimens from the lesional and perilesional skin of five patients with severe AD as well as from the normal skin of five healthy human controls.Real-time fluorescence-based reverse transcription (RT)-PCR was employed to measure the mRNA expression levels of miR-155,RORγt and IL-17 in peripheral blood mononuclear cells (PBMCs) and skin specimens,flow cytometry to detect the percentage of Th17 cells in PBMCs,enzyme-linked immunosorbent assay (ELISA) to determine the plasma concentration of IL-17.Statistical analysis was done using independent sample's t test,one-way analysis of variance followed by the least significant difference test,and linear correlation analysis.Results Compared with the healthy controls,the patients with AD showed a significant increase in Th17 cell percentage (1.78% ± 0.52% vs.0.47% ± 0.15%,P< 0.01),mRNA expression levels of miR-155 (5.78 ± 1.78 vs.1.82 ± 0.46,P< 0.01),RORγt (6.08 ± 1.04 vs.1.64 ± 0.52,P< 0.01) and IL-17 (7.09 ± 1.75 vs.1.71 ± 0.46,P< 0.01),as well as in the plasma concentration of IL-17 ((2.51 ± 6.15) pg/ml vs.(11.80 ± 2.24) pg/ml,P< 0.01).There was a sequential decrease in the expression levels of miR-155,RORγt and IL-17 mRNA from lesional skin,perilesional skin to normal skin (F =41.803,17.040 and 37.064 respectively,all P < 0.01).The miR-155 mRNA expression level in PBMCs was positively correlated with the SCORing Atopic Dermatitis (SCORAD) index,Th17 cell percentage,RORγt and IL-17 mRNA expression levels as well as IL-17 plasma concentration (r =0.405,0.426,0.402,0.410 and 0.408 respectively,all P < 0.05).Similarly,the miR-155 expression level was positively correlated with RORγt and IL-17 mRNA expression levels in lesional and paralesional specimens (r =0.428 and 0.435 respectively,both P < 0.05).Conclusion The up-regulated expression of miR-155,Th17 cells and their effector cytokine IL-17 may be associated with the development of AD.
10.Expression of CK19 in sentinel lymph node for breast cancer and its clinical significance
Chunmei YE ; Fengling LUO ; Mingxing XUE ; Bo CHEN ; Ziming HUANG ; Song ZHAO ; Rong WANG
Clinical Medicine of China 2010;26(7):751-753
Objective To detect the expression of CK19 in sentinel lymph node for breast cancer and to evaluate its clinical significance.Methods SLNs of 30 patients with breast cancer were successfully detected with methylene blue,both SLN and ALN were examined by hematoxylin and eosin staining ( HK staining). The expression level of CK19 in SLNs were assessed by RT-PCR and Western Blot for mRNA and Protein respectively. Results Detection positive rate of SLN melastases was 26. 67% (8/30) by HE staining,and 1 case of false negative. Through RT-PCR,12 cases,including all the positive cases detected by routine pathological examination,were detected to be positive in the expression of CK19mRNA, with a positive rate of 40. 0% ( 12/30) , and a positive rate of 36. 67% (11/30) for CK19 was observed by Western Blot There was a significant difference between RT-PCR or Western Blot methods and routine pathological examination ( P < 0.05 ) for the detection efficiency of SI.Ns melastases. Conclusions CK19rnRNA may be a potential tumor marker for detecting micrometastasis in SLNs of breast cancer. The combined utilization of locating SLNs and detecting CK19 mRNA expression by RT-PCR instead of histopathological examination can greatly enhance the detection efficiency of SLN micrometastasis of breast cancer.