1.Cloning and functional characterization of a cDNA encoding isopentenyl diphosphate isomerase involved in taxol biosynthesis in Taxus media.
Tian SHEN ; Fei QIU ; Min CHEN ; Xiaozhong LAN ; Zhihua LIAO
Acta Pharmaceutica Sinica 2015;50(5):621-6
Taxol is one of the most potent anti-cancer agents, which is extracted from the plants of Taxus species. Isopentenyl diphosphate isomerase (IPI) catalyzes the reversible transformation between IPP and DMAPP, both of which are the general 5-carbon precursors for taxol biosynthesis. In the present study, a new gene encoding IPI was cloned from Taxus media (namely TmIPI with the GenBank Accession Number KP970677) for the first time. The full-length cDNA of TmIPI was 1 232 bps encoding a polypeptide with 233 amino acids, in which the conserved domain Nudix was found. Bioinformatic analysis indicated that the sequence of TmIPI was highly similar to those of other plant IPI proteins, and the phylogenetic analysis showed that there were two clades of plant IPI proteins, including IPIs of angiosperm plants and IPIs of gymnosperm plants. TmIPI belonged to the clade of gymnosperm plant IPIs, and this was consistent with the fact that Taxus media is a plant species of gymnosperm. Southern blotting analysis demonstrated that there was a gene family of IPI in Taxus media. Finally, functional verification was applied to identify the function of TmIPI. The results showed that biosynthesis of β-carotenoid was enhanced by overexpressing TmIPI in the engineered E. coli strain, and this suggested that TmIPI might be a key gene involved in isoprenoid/terpenoid biosynthesis.
2.Efficacy of tepronone and folic acid in the treatment of chronic atrophic gastritis evaluated by the marking targeting biopsy
Lifeng QIU ; Lan ZHAO ; Yanfei FANG ; Shujie CHEN ; Jianmin SI
Chinese Journal of Digestion 2013;33(4):231-234
Objective To explore the efficacy of tepronone and folic acid in the treatment of chronic atrophic gastritis (CAG) evaluated by the marking targeting biopsy (MTB).Methods A total of 224 H.pylori negative CAG patients were selected and divided into group A (n 96,tepronone 50 mg/time,folic acid 10 mg/time,three times/day),group B (n=23,tepronone 50 mg/time,three times/day),group C (n=74,unspecific treatment) and group D (n=31,no treatment).The treatment course lasted for one year.The clinical symptoms improvement of each group was observed before and after treatment.The pathological improvement of gastric mucosa by MTB was inspected before and after treatment.The chi square test was performed for the comparison between groups.Results The total efficacy rates of group A,B,C and D were 43.8% (42/96),39.1% (9/23),33.8%(25/74) and 32.3% (10/31) respectively,there was no significant difference between groups (x2 =2.328,P =0.507).For the significant efficacy rate of gastric mucosa pathological improvement,group A was compared with group D,group A was compared with group C and group B was comparedwith group D,the differences were significant (x2 =14.520,14.628 and 8.995,all P<0.01).In the total efficacy rate of gastric mucosa pathological improvement,group A (49.8%,131/263) was compared with group D (24.2%,16/66),group A was compared with group C (35.9%,66/184)and group B (44.7%,21/47) was compared with group D,the differences were significant (x2 =13.953,8.535 and 5.207,all P<0.05).Conclusion Teprenone alone or teprenone and folic acid combination can obviously improve pathological changes of CAG patients.
3.Expression of acid sensing ion channel 3 in the lung tissue of rats with, acute lung injury
Lan SHAO ; Yansheng CHEN ; Shaoqun XU ; Haibo QIU
Chinese Journal of Emergency Medicine 2009;18(5):466-470
Objective To explore the expression of acid seining ion channels-3 (ASIC3) in lung tissue in rats with acute lung injury (ALI). Method Twenty four male Sprague-Dawley (SD) rats were randomly divided into four groups: LPS groups (LPS 2 h, LPS 4 h, LPS 6 h group, n=6), stimulated by LPS for 2, 4, 6 hours, respectively; normal control group, injected with saline (NS group, n=6). The ALI models were produced through venous injection of LPS, and the criteria was the characteristic pathological changes in the lung tissue. Ar-terial blood gas analysis was observed, lung wet and dry weight ratio (W/D), lung histopathology and ASIC3 ex-pression were detected. Data were expressed as mean±standard deviation. Independent Sample T test and One-way ANOVA and Kendall's tau_b were used for comparison in SPSS 13.0, and changes were considered as statistieal-ly significant if P value was less than 0.05. Results The partial pressure of oxygen (PaO2) in LPS 2 h, LPS 4 h, LPS 6 h group was (67.47±6.01), (59.17±7.18), (52.54±7.62) , respecively, and was significantly lower than that in eontrol group (98.15±1.06) (P<0.01). Compare with control group, pH was significantly lower in LPS4 h group (7.28±0.04), LPS6 h group (7.24±0.03) (P<0.01). Inflammation cells gradually increased, alveolar septum was widened, edema existed in interstitial spaces, and pulmonary structures gradually destroyed in LPS groups.The expression of ASIC3 in LPS4 h, LPS 6 h group was (205.91±10.12), (196.51± 18.60), respectively, and was significantly lower thanthat in control group (220.23±10.11) (P<0.05). The W/D in LPS 6 h group was (5.18±0.21), and was significantly higher than that in the control group (4.45± 0.18) (P<0.05). Conclusions ASIC3 is expressed in alveolar epithelial cells and bronchial epithehal cells in LPS-induced ALI rats.
5.Mapping of the B Cell Neutralizing Epitopes on ED III of Envelope Protein from Dengue Virus.
Yaying LIN ; Kun WEN ; Yonghui GUO ; Liwen QIU ; Yuxian PAN ; Lan YU ; Biao DI ; Yue CHEN
Chinese Journal of Virology 2015;31(6):665-673
Dengue virus (DENV) envelope [E] protein is the major surface protein of the virions that indued neutralizing antibodies. The domain III of envelope protein (EDIII) is an immunogenic region that holds potential for the development of vaccines; however, the epitopes of DENV EDIII, especially neutralizing B-cell linear epitopes, have not been comprehensively mapped. We mapped neutralizing B-cell linear epitopes on DENV-1 EDIII using 27 monoclonal antibodies against DENV-1 EDIII proteins from mice immunized with the DENV-1 EDIII. Epitope recognition analysis was performed using two set of sequential overlapping peptides (16m and 12m) that spanned the entire EDIII protein from DENV-1, respectively. This strategy identified a DENV-1 type- specific and a group-specific neutralizing epitope, which were highly conserved among isolates of DENV-1 and the four DENV serotypes and located at two regions from DENV-1 E, namely amino acid residues 309-320 and 381-392(aa 309-320 and 381-392), respectively. aa310 -319(310KEVAETQHGT319)was similar among the four DENV serotypes and contact residues on aa 309 -320 from E protein were defined and found that substitution of residues E309 , V312, A313 and V320 in DENV-2, -3, -4 isolates were antigenically silent. We also identified a DENV-1 type-specific strain-restricted neutralizing epitope, which was located at the region from DENV-1 E, namely amino acid residues 329-348 . These novel type- and group-specific B-cell epitopes of DENV EDIII may aid help us elucidate the dengue pathogenesis and accelerate vaccine design.
Amino Acid Sequence
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Animals
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Antibodies, Neutralizing
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immunology
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Dengue
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virology
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Dengue Virus
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chemistry
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genetics
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immunology
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Epitope Mapping
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Epitopes, B-Lymphocyte
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chemistry
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genetics
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immunology
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Humans
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Mice
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Molecular Sequence Data
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Viral Envelope Proteins
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chemistry
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genetics
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immunology
6.Effects of benazepril on cardiac function, free oxygen radicals, sarcoplasmic reticulum Ca~(2+)- ATPase following cardiac ischemia- reperfusion in spontaneously hypertensive rats
Yesong WANG ; Hong MA ; Jianwen CHEN ; Yuan HU ; Xiujian LAN ; Fanhua QIU
Chinese Journal of Pathophysiology 1986;0(03):-
AIM: To investigate the effects of angiotensin converting enzyme inhibitor (ACEI), benazepril(B), on cardiac function, free oxygen radicals, sarcoplasmic reticulum(SR) Ca~(2+)-ATPase following ischemia-reper-fusion in sportaneously hypertensive rats (SHRs). METHODS: Thirty 10-week-old female SHRs were randomly assigned into two groups: group SHR was control; The animal in group SHR+B was given with 10 mg/kg of benazepril perday. Another 15 Wistar rats with the same age and sex were normal control (group Wistar). After 12 weeks of pretreatment, all rats in each group were subjected to 30 min of left anterior descending coronary artery occlusion and 30 min of reperfusion. Hemodynamic parameters, left heart-to-body weight ratio(LVW/BW), myocardial malondialdehyde (MDA) concentration, superoxide dismutase (SOD) activity, and SR Ca~(2+)-ATPase activity were measured. RESULTS: Compared to group Wistar, the rats in group SHR had higher blood pressure, LVW/BW and myocardial MDA concentration, more serious left cardiac function injury and lower myocardial SOD activity and SR Ca~(2+)-ATPase activity; group SHR+B had lower myocardial MDA concentration, higher myocardial SOD activity, but no difference in blood pressure, LVW/BW, the degree of left cardiac function injury and myocardial SR Ca~(2+)-ATPase activity. CONCLUSION: Benazepril can attenuate ischemia-reperfusion-induced cardiac function injury by regression of left ventricular hypertrophy (LVH), improving SR Ca~(2+)-ATPase activity and decreasing oxygen free radicals injury in SHRs.
7.The performance of pediatric risk of mortality score in pediatric patients with severe hand,foot and mouth disease
Lan LUO ; Caixia LONG ; Peng CHEN ; Xiulan LU ; Zhenghui XIAO ; Xiao LIU ; Chao ZUO ; Jun QIU
Chinese Pediatric Emergency Medicine 2015;22(8):567-570
Objective To comment the severity of severe hand,foot and mouth disease(HFMD)by pediatric risk of mortality score(PRISM),and assess the performance of PRISM in predicting mortality or complication probability in HFMD.Methods Four hundred and twenty-four severe HFMD pediatric patients were recruited in the study from 1th Jan 2010 to 31th June 2013.Information on the outcome and the varia-bles required to calculate PRISM score were collected.The logistic regression model developed in the learning sample was evaluated in the test sample by calculating the area under the receiver operating characteristic (ROC)curve to assess discrimination pneumorrhagia and death.Calibration across deciles of risk was evalua-ted using the Hosmer-Lemeshow goodness-of-fit χ2 test.Results The area under the ROC curve were 0.87 (95%CI 0.80~0.94 )for PRISM in predicting pneumorrhagia probability.The area under the ROC curve were 0.87(95%CI 0.80~0.95)for PRISM in predicting mortality probability.The PRISM in observed and expected pneumorrhagia did not demonstrate good calibration at ten mortality risk intervals (χ2 =36.66, P<0.001 ).The PRISM in observed and expected mortality did not demonstrate good calibration at ten mortali-ty risk intervals(χ2 =41.11,P<0.001).Conclusion The PRISM score is demonstrated good discrimination of pneumorrhagia and death in HFMD pediatric patients,but the performance of calibration is not good.
8.A 2-Year Follow-up Study of Residual Hearing after Cochlear Implantation
Sumei QIU ; Lin LIN ; Ping ZHOU ; Xiaobing LAN ; Minghui ZHENG ; Huijuan CHEN
Journal of Audiology and Speech Pathology 2013;(5):523-525
Objective To investigate the effects of cochlear implantation on residual hearing and to evaluate the potential impact of long -term electrical stimulations on residual hearing .Methods 58 hearing impaired children with cochlear implants were included in this study .All subjects could cooperate with behavioral audiometry .Audio-metric evaluations were carried out pre -implantation and 3 ,12 ,24 months post -implantation respectively .Of 58 subjects ,43 were followed up more than 1 year and 17 were followed up more than 2 years .Results All 58 subjects showed significant differences (P<0 .05) between pre- and 3 months post-implantation of residual hearing at the individual frequencies of 0 .25 ,0 .5 ,1 ,2 and 4 kHz .43 subjects followed up more than 1 year showed statistic differences (P<0 .05) between pre- and 3 months post -implantation ,pre- and 12 months post-implantation at 0 .25 ,0 .5 ,1 ,2 and 4 kHz respectively .Comparing 3 months with 12 months post -implantation ,there were sta-tistic differences at 0 .25 and 0 .5 kHz ,while no significant difference (P>0 .05) at 1 ,2 and 4 kHz .Of 17 subjects followed up more than 2 years ,there were significant differences (P<0 .05) between pre- and various return visits post-implantation .Post-implantation return visits ,there were significant differences between 3 months and 12 , 24 months at 0 .25 and 0 .5 kHz respectively ,not any significant differences on 1 ,2 and 4 kHz .There were no sig-nificant differences on each frequency between 12 months and 24 months post- implantation .Conclusion Residual hearing after cochlear implantation could decrease to some extent for various reasons .There were significant differ-ences between 3 and 12 months post-implantation at 0 .25 and 0 .5 kHz .Not any significant differences were ob-served between 12 months and 24 months post-implantation at each frequency .
9.Genetic diagnosis of a patient with non-syndromic variants of congenital neutropenia
Shengli XUE ; Yan CHEN ; Qiaocheng QIU ; Yufeng FENG ; Lan DAI ; Man QIAO ; Depei WU
Chinese Journal of Internal Medicine 2011;50(11):922-925
ObjectiveTo explore the procedures and methods for genetic diagnosis in one nonsyndromic variants of congenital neutropenia (NSVCN) patient and its pathogenic mutation.Methods Genomic DNA was prepared from one NSVCN patient who had progressed to chronic myelomonocytic leukemia and ELA2,HAX1,WASp and GFI1 genes were amplified and sequenced.Results A novel compound heterogeneous mutation consisting of two frame-shift mutations (c.430-1insG and c.655- 9del5bp) was found in HAX1 gene.ConclusionA practically genetic diagnosis procedure for NSVCN has been established,and the novel HAX1 gene mutation may contribute to the etiology of NSVCN.
10.Cloning and functional characterization of a cDNA encoding isopentenyl diphosphate isomerase involved in taxol biosynthesis in Taxus media.
Tian SHEN ; Fei QIU ; Min CHEN ; Xiao-zhong LAN ; Zhi-hua LIAO
Acta Pharmaceutica Sinica 2015;50(5):621-626
Taxol is one of the most potent anti-cancer agents, which is extracted from the plants of Taxus species. Isopentenyl diphosphate isomerase (IPI) catalyzes the reversible transformation between IPP and DMAPP, both of which are the general 5-carbon precursors for taxol biosynthesis. In the present study, a new gene encoding IPI was cloned from Taxus media (namely TmIPI with the GenBank Accession Number KP970677) for the first time. The full-length cDNA of TmIPI was 1 232 bps encoding a polypeptide with 233 amino acids, in which the conserved domain Nudix was found. Bioinformatic analysis indicated that the sequence of TmIPI was highly similar to those of other plant IPI proteins, and the phylogenetic analysis showed that there were two clades of plant IPI proteins, including IPIs of angiosperm plants and IPIs of gymnosperm plants. TmIPI belonged to the clade of gymnosperm plant IPIs, and this was consistent with the fact that Taxus media is a plant species of gymnosperm. Southern blotting analysis demonstrated that there was a gene family of IPI in Taxus media. Finally, functional verification was applied to identify the function of TmIPI. The results showed that biosynthesis of β-carotenoid was enhanced by overexpressing TmIPI in the engineered E. coli strain, and this suggested that TmIPI might be a key gene involved in isoprenoid/terpenoid biosynthesis.
Amino Acid Sequence
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Carbon-Carbon Double Bond Isomerases
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genetics
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Cloning, Molecular
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DNA, Complementary
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genetics
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Escherichia coli
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Paclitaxel
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biosynthesis
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Phylogeny
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Plant Proteins
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genetics
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Taxus
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enzymology
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genetics