1.Advances of the matrix assisted laser desorption ionization-time of flight-mass spectrometry in lung cancer
Journal of International Oncology 2014;41(5):344-347
Matrix-assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS) is a kind of the latest mass spectrometry,which has many advantages including high throughput,rapidity,excellent repeatability and high sensitivity.It has been an important tool for both the research and clinical application in lung cancer and has an optimistic prospect in the many fields,such as early diagnosis,screening and evaluation of therapeutic effect.
2.Regulatory effects of miR-146a on Cryptococcus neoformans-induced inflammation factors production in THP-1 cells
Zhijun HAN ; Hongyu HUANG ; Min ZHANG ; Yunyu WEI ; Jingbo LIU ; Zihe YAN
Chinese Journal of Microbiology and Immunology 2013;(10):740-743
Objective To investigate the regulatory effects of miR-146a on inflammation factors production in C ryptococcus neoformans treated THP-1 cells.Methods Cryptcoo ccus neoformans strains were heat killed .Fluorescence quantitative RTP-CR and ELISA were used to detect the levels of IL -1βand TNF-αin THP-1 cells treated with heat-killed Cryptococcus neoformans.In addition, the production of IL-1βand TNF-αwere analyzed before and after Dectin-1or TLR 4 blocked .THP-1 cell lines that were respectively transfected with miR-146 a mimics and inhibitors were constructed and the production of IL-1βand TNF-αin these cell lines were analyzed after interfered with Cryptococcus neoformans.Results With the interference of heat-killed Cryptococcus neoformans, the expression of miR-146a was up-regulated in THP-1 cells, but was down-regulated after Dectin-1 or TLR4 blocked.The expressions of IL-1βand TNF-αinduced by heat-killed Cryptococcus neoformans were enhanced in miR-146a mimics transfected THP-1 cells, but was inhibi-ted in inhibitors transfected THP-1 cells.Conclusion Heat-killed Cryptococcus neoformans could up-regu-late miR-146a expression in THP-1 cells via Dectin-1 and TLR.miR-146a could negatively regulate the ex-pressions of IL-1βand TNF-αinduced by Cryptococcus neoformans.
3.MiR-146a regulate the proliferation and interleukin-2 production of T helpers from primary biliary cirrhosis patients
Zhijun HAN ; Zhide HU ; Jingbo LIU ; Zihe YAN ; Tianli REN ; Anmei DENG ; Renqian ZHONG
Chinese Journal of Rheumatology 2012;16(8):541-544
Objective To investigate the effect of miR-146a on the proliferation and interleukin (IL)-2 production of T helper cells from primary biliary cirrhosis (PBC) patients.Methods MiR-146a in the peripheral blood mononuclear cells (PBMC),monocytes,T helper cells,cytotoxic T cells and B cells from 20 confirmede PBC patients and age/sex matched healthy controls were detected by quantitative PCR.By gainand-loss of function,the miR-146a's effect on anti-CD3/anti-CD28 activated T helper's proliferation and IL-2 production ability were measured by CCK-8 approach and enzyme linked immunosorbent assays (ELISA),respectively.Statistical analysis were carried out by t-test.Results PBMCs (0.46±0.20 vs 1.00±0.26; t=7.47,P<0.01),T helpers (0.33±0.13 vs 1.00±0.14; t=6.15,P<0.01) and monocytes (0.56±0.11 vs 1.00±0.11; t=4.97,P<0.05),but not B cells (0.91±0.06 vs 1.00±0.14; t=0.97,P>0.05) and cytotoxic T cells (0.98±0.15vs 1.00±0.12; t=0.22; P>0.05) from PBC patients had lower miR-146a expression level than that of healthy controls.Inducible up expression of miR-146a was observed in PBC patients'T helpers stimulated with antiCD3/anti-CD28 (1.00±0.18 vs 9.12±2.05; t=8.81; P<0.01).The activated T helpers from PBC patients had higher proliferative ability [PBC:0.35±0.06 (A); healthy controls:0.26±0.04 (A); t=2.83; P<0.05] and increased IL-2 production [PBC: (685.60±109.19 pg/ml)]; Healthy controls: [(512.20±72.26) pg/ml; t=2.96; P<0.05 ] than those of healthy controls.For activated T helpers,the proliferation ability,as well as IL-2 production,was enhanced by miR-146a.Conclusion MiR-146a can down regulate the proliferation and IL-2 releasing of activated T helpers.The reduced miR-146a expression enhances IL-2 production and promotes proliferation of T helper of PBC patients,thus,may be involved in the pathogenesis of PBC.
4.The ERP brain topographic map study on mental rotation of depressions
Jiu CHEN ; Laiqi YANG ; Guangxiong LIU ; Xingqu WU ; Yan ZHANG ; Wentao MA ; Zihe DENG
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(2):135-138
ObjectiveTo explore the change of the event related potential brain topographic map on depression' mental rotation,and to perfect the brain function relation map for depression in space ability.Methods 32 depression and 29 normal healthy people were tested to make mental rotation tasks in the brain ERP system.The distribution of the changing brain topographic map were observed.Results ( 1 ) Compared with the control group ( error rate ( 29±9 ) %,response time ( 604.74 ± 54.39 ) ms,the error rate was significantly higher and response time was significantly longer in depression (error rate( 33 ± 15 )%,response time(755.22 ± 70.18 )ms,P<0.05).(2) Compared with the control group (N100:PZ( -3.78 ± 1.05)μV,CZ( -5.67 ±2.21)μV,P3( -2.34 ±0.59) μV,P4( -2.92 ±0.80) μV ;P500:PZ(7.35 ±2.61 ) μV,CZ(7.65 ± 2.42) μV,P3 (6.53 ±2.11 ) μV,P4 ( 7.29 ± 2.57 ) μV ),the total volatility was significantly lower in depression ( N 100:PZ ( - 0.31 ±0.09)μV,CZ( -2.27 ±0.57)μV,P3( -0.30 ±0.07) μV,P4( -0.33 ±0.08) μV;P500:PZ(6.04 ±2.16)μV,CZ ( 5.92 ± 2.01 ) μV,P3 ( 6.02 ± 2.11 ) μV,P4 (6.01 ± 2.34 ) μV,P < 0.05 ) and the excitability difference of the left and right parietal-occipital lobe was disappeared (P>0.05) ; Compared with the control group,in N100 the normal and mirror excitability was significantly lower,and in P500 the normal excitability was significantly lower,but mirror was significantly higher in depression (P < 0.05 ).Compared with the left and right brain,the normal excitability in the right parietal-occipital lobe was significant higher (P < 0.05 ),but the mirror excitement difference was disappeared in depression (P> 0.05 ),and the normal and mirror excitement in the right parietal-occipital lobe was both significantly higher in normal healthy people (P < 0.05 ).ConclusionDepressed patients; mental rotation ability is impaired.And the negative potential for looking forward to reaction is lower and exist the right advantage hemisphere brain in normal,but mirror advantage hemisphere disappears in depressed patients.This study suggests the brain topographic map of mental rotation ability damaged can be used as the clinical auxiliary diagnosis index.
5.Increased expression of microRNA-146a in peripheral blood mononuclear cells of patients with chronic immune thrombocytopenic purpura and its clinical significance
Zhijun HAN ; Zhide HU ; Anmei DENG ; Yi SUN ; Jingbo LIU ; Yuanlan HUANG ; Zihe YAN ; Renqian ZHONG
Chinese Journal of Microbiology and Immunology 2011;31(1):81-84
Objective To investigate the increased expression of microRNA-146a(miR-146a) in peripheral blood mononuclear cells (PBMC) of patients with chronic immune thrombocytopenic purpura (ITP) and its clinical significance. Methods Twenty-eight patients with chronic ITP and 28 healthy controls matched with age and gender were enrolled in this study. Fluorescent quantitative PCR reaction was used to detect the relative expression of miR-146a in their PBMC. The serum concentration of TNF-α, IL-2,IL-1 β and IFN-γ were measured by ELISA. CCK-8 method was used to detect the proliferation ability of PBMC , which transfected with miR-146a mimics or inhibitor and then stimulated with platelet . Results The relative expression of miR-146a in ITP patients was higher than that of healthy controls. The increased expression of miR-146a was negatively correlated with the serum TNF-α, IL-2 and IFN-γ. The PBMC transfected with miR-146a mimics had reduced expression of IL-2 and proliferation when stimulated with platelet.In contrast, the opposite effect was observed with the miR-146a inhibitors transfection. Conclusion MiR146a was involved in the pathogenesis of chronic ITP by controlling IL-2 production and PBMC proliferation.Thus, it may be a potential therapy target for chronic ITP.
6.Experiment study of the acellular bovine pericardium treated by dye-mediated photooxidation used as engineering heart tissue
Zhenliang ZHANG ; Jianye ZHOU ; Shengshou HU ; Liqun LIU ; Pingping SUN ; Zihe YANG ; Jut LI
Chinese Journal of Thoracic and Cardiovascular Surgery 2011;27(8):485-488
ObjectiveTo evaluate the feasibility of constructing tissue engineering cardiac patch with photooxidationfixed acellular bovine pericardium.MethodsFresh bovine pericardia were treated by dye-mediated photooxidation after decellularization.Some of them were seeded with bone marrow stromal cells(MSCs) isolated from male SD rats to construct cardiac patches.Myocardial infarction(MI) model was made in female SD rats by left anterior descending coronary ligation(LAD).One week later, the confirmed MI rats were divided into three groups randomly, group MI (n = 15)without any treatment; group P (n = 18) with photooxidated pericardia implantation ; group P + C (n = 18) with seeded pericardia implantation.A sham group (n = 10) was also performed with opening and closing chest twice only.The heart were explanted at 2 or 4 weeks after implantation, and examined histologically and immunohistochemically.The heart function was evaluated by echocardiography at 4 weeks before excising the rats.ResultsThere were no cells or cell debris remained in bovine pericardium tissue.The fiber structure became condensed after photooxidation.The seeded cells formed a continuous layer on the surface of the tissue.The pericardial degradation level and newly formed microvessel density were larger in group P + C than in group P after 2 [ (13.7 ±5.2)个/mm2 vs (7.1 ±3.1)个/mm2, P<0.05]and4 [(22.6 ±4.9)个/mrn2 vs (14.1 ±5.3)个/mm2, P<0.05]weeks.Four weeks after transplantation, cardiac echocardiography showed left ventricular ejection fraction(LVEF) was lower in group MI (44.8 ± 4.4) % and group P (48.4 ± 5.0) % compared with group P + C (49.3 ± 4.8) %, left ventricular fractional shorterning(LVFS) was lower in group MI (18.0 ± 2.2) % and group P (19.8 ± 2.5) % compared with group P + C (20.4 ±2.5) %, the difference between P + C and MI was significant.ConclusionTransplantation of the tissue engineered bovine pericardial patches with dye-mediated photooxidation can improve heart function in MI rats.This kind of patches demonstrates a promising prospect in the future.
7.Event-related potentials and the mirror-normal differences of object rotation in first-episode schizophrenia
Jiu CHEN ; Laiqi YANG ; Yongxiong CHEN ; Lanlan LI ; Guangxiong LIU ; Yan ZHANG ; Xingqu WU ; Wentao MA ; Zihe DENG
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(3):209-211
Objective To explore the brain electrophysiological mechanism of object rotation in first-episode schizophrenia.Methods 30 patients with schizophrenia and 28 normal healthy people,who were from the Center for Mental Disease Control and Prevention,Third Hospital of PLA,took part in the mental rotation tasks,then the incubation period and amplitude of P500,and the wrong number and reaction time were measured.Results Compared with control group ( normal:(494.16 ± 34.68 ) ms,( 9.56 ± 2.54) μV; mirror:(496.51 ± 33.10 ) ms,(6.38 ± 2.41 ) μV),schizophrenia' incubation periods were significantly delayed ( normal:( 571.30 ± 51.21 ) ms;mirror:(573.41 ±39.27) ms) and volatility were significantly lower ( normal:(4.26 ± 1.01 ) μV; mirror:(3.61± 1.21 )μV) in normal and mirror rotation (P<0.05 ).The mirror-normal differences were not significant on the incubation periods of two groups (P > 0.05 ) ; the mirror-normal image differences were not significant on the patient group' volatility (P > 0.05 ) ; the normal volatility was significantly higher than mirror in the control group (P < 0.05).Conclusion Schizophrenia'mental rotation ability is impaired,and mirror-normal differences on mental rotation are disappeared.It can be used as an early-stage clinical auxiliary diagnosis index.
8.Crystal structure of a secreted lipase from Gibberella zeae reveals a novel "double-lock" mechanism.
Zhiyong LOU ; Ming LI ; Yuna SUN ; Ye LIU ; Zheng LIU ; Wenping WU ; Zihe RAO
Protein & Cell 2010;1(8):760-770
Fusarium graminearum (sexual stage: Gibberella zeae) is the causative agent of Fusarium Head Blight (FHB), which is one of the most destructive plant disease of cereals, accounting for high grain yield losses, especially for wheat and maize. Like other fungal pathogens, several extracellular enzymes secreted by G. zeae are known to be involved in host infection. Among these secreted lipases, G. zeae lipase (GZEL), which is encoded by the FGL1 gene, was demonstrated to be crucial to G. zeae pathogenicity. However, the precise mechanism of GZEL remains unclear due to a lack of detailed structural information. In this study, we report the crystal structure of GZEL at the atomic level. The structure of GZEL displays distinct structural differences compared to reported homologues and indicates a unique "double lock" enzymatic mechanism. To gain insight into substrate/inhibitor recognition, we proposed a model of GZEL in complex with substrate and the lipase inhibitor ebelactone B (based on the reported structures of GZEL homologues), which defines possible substrate binding sites within the catalytic cleft and suggests an "anti sn-l" binding mode. These results pave the way to elucidating the mechanism of GZEL and thus provide clues for the design of anti-FHB inhibitors.
Amino Acid Sequence
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Catalytic Domain
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Crystallography, X-Ray
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Gibberella
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enzymology
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Lactones
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chemistry
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Lipase
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chemistry
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metabolism
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Models, Molecular
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Molecular Sequence Data
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Oleic Acid
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chemistry
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Protein Binding
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Protein Structure, Secondary
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Protein Structure, Tertiary
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Sequence Alignment
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Sequence Homology, Amino Acid
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Substrate Specificity
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Surface Properties
9.Structure analysis of the extracellular domain reveals disulfide bond forming-protein properties of Mycobacterium tuberculosis Rv2969c.
Lu WANG ; Jun LI ; Xiangxi WANG ; Wu LIU ; Xuejun C ZHANG ; Xuemei LI ; Zihe RAO
Protein & Cell 2013;4(8):628-640
Disulfide bond-forming (Dsb) protein is a bacterial periplasmic protein that is essential for the correct folding and disulfide bond formation of secreted or cell wallassociated proteins. DsbA introduces disulfide bonds into folding proteins, and is re-oxidized through interaction with its redox partner DsbB. Mycobacterium tuberculosis, a Gram-positive bacterium, expresses a DsbA-like protein ( Rv2969c), an extracellular protein that has its Nterminus anchored in the cell membrane. Since Rv2969c is an essential gene, crucial for disulfide bond formation, research of DsbA may provide a target of a new class of anti-bacterial drugs for treatment of M.tuberculosis infection. In the present work, the crystal structures of the extracellular region of Rv2969c (Mtb DsbA) were determined in both its reduced and oxidized states. The overall structure of Mtb DsbA can be divided into two domains: a classical thioredoxin-like domain with a typical CXXC active site, and an α-helical domain. It largely resembles its Escherichia coli homologue EcDsbA, however, it possesses a truncated binding groove; in addition, its active site is surrounded by an acidic, rather than hydrophobic surface. In our oxidoreductase activity assay, Mtb DsbA exhibited a different substrate specificity when compared to EcDsbA. Moreover, structural analysis revealed a second disulfide bond in Mtb DsbA, which is rare in the previously reported DsbA structures, and is assumed to contribute to the overall stability of Mtb DsbA. To investigate the disulphide formation pathway in M.tuberculosis, we modeled Mtb Vitamin K epoxide reductase (Mtb VKOR), a binding partner of Mtb DsbA, to Mtb DsbA.
Amino Acid Sequence
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Bacterial Proteins
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chemistry
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metabolism
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Catalytic Domain
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Crystallography, X-Ray
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Disulfides
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chemistry
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Escherichia coli
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metabolism
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Escherichia coli Proteins
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chemistry
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metabolism
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Molecular Docking Simulation
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Molecular Sequence Data
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Mycobacterium tuberculosis
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metabolism
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Oxidation-Reduction
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Protein Disulfide-Isomerases
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chemistry
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metabolism
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Protein Folding
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Protein Structure, Tertiary
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Sequence Alignment
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Static Electricity
10.Insight into the Ebola virus nucleocapsid assembly mechanism: crystal structure of Ebola virus nucleoprotein core domain at 1.8 Å resolution.
Shishang DONG ; Peng YANG ; Guobang LI ; Baocheng LIU ; Wenming WANG ; Xiang LIU ; Boran XIA ; Cheng YANG ; Zhiyong LOU ; Yu GUO ; Zihe RAO
Protein & Cell 2015;6(5):351-362
Ebola virus (EBOV) is a key member of Filoviridae family and causes severe human infectious diseases with high morbidity and mortality. As a typical negative-sense single-stranded RNA (-ssRNA) viruses, EBOV possess a nucleocapsid protein (NP) to facilitate genomic RNA encapsidation to form viral ribonucleoprotein complex (RNP) together with genome RNA and polymerase, which plays the most essential role in virus proliferation cycle. However, the mechanism of EBOV RNP formation remains unclear. In this work, we solved the high resolution structure of core domain of EBOV NP. The polypeptide of EBOV NP core domain (NP(core)) possesses an N-lobe and C-lobe to clamp a RNA binding groove, presenting similarities with the structures of the other reported viral NPs encoded by the members from Mononegavirales order. Most strikingly, a hydrophobic pocket at the surface of the C-lobe is occupied by an α-helix of EBOV NP(core) itself, which is highly conserved among filoviridae family. Combined with other biochemical and biophysical evidences, our results provides great potential for understanding the mechanism underlying EBOV RNP formation via the mobility of EBOV NP element and enables the development of antiviral therapies targeting EBOV RNP formation.
Crystallography, X-Ray
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Ebolavirus
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physiology
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Humans
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Nucleoproteins
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chemistry
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genetics
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metabolism
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Protein Structure, Tertiary
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Structure-Activity Relationship
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Virus Assembly
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physiology