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Shizhen Wang,Jing Wang,Xiaofen Zhou,Yingxia Guo,Baishan Fang 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.4
The substitution of the catalytic zinc ion of glycerol dehydrogenase (GDH) from Klebsiella pneumonia sp. by divalent metal ions, Mn2+ and Mg2+, enabled improvements of activity, substrate promiscuity and stability. The activity of Mn-GDH and Mg-GDH improved several folds in comparison to the native GDH. The activity of substituted GDH towards non-natural substrates, 4-chloroacetoacetate,3-chloroacetylpyridine, p-chloroacetophenone, and acetophenone was 30 folds higher than native GDH. Manganese substitution increased the half-life of GDH by 6 folds at 60and 70oC. The two-fraction first order inactivation models fitted the nonlinear thermal inactivation curves well. Combined with the kinetic and thermodynamic analysis,further mechanistic insights to the metal ion roles in thermostability enhancements were studied. The thermodynamic parameters of inactivation, enthalpy, entropy and the Gibbs free energy indicated that Mn-GDH was stabilized entropically and elucidated the mechanisms of enzyme inactivation.
Wang, Xinglong,Liu, Li,Liu, Sixiu,Sun, Xiaoqing,Deng, Zhongxiang,Pi, Yan,Sun, Xiaofen,Tang, Kexuan Korean Society for Biochemistry and Molecular Biol 2004 Journal of biochemistry and molecular biology Vol.37 No.5
A new CRT binding factor (CBF) gene designated Cbcbf25 was cloned from Capsella bursa-pastoris, a wild grass, by the rapid amplification of cDNA ends (RACE). The full-length cDNA of Cbcbf25 was 898 bp with a 669 bp open reading frame (ORF) encoding a putative DRE/CRT (LTRE)-binding protein of 223 amino acids. The predicted CbCBF25 protein contained a potential nuclear localization signal (NLS) in its N-terminal region followed by an AP2 DNA-binding motif and a possible acidic activation domain in the C-terminal region. Bioinformatic analysis revealed that Cbcbf25 has a high level of similarity with other CBF genes like cbf1, cbf2, and cbf3 from Arabidopsis thaliana, and Bncbf5, Bncbf7, Bncbf16, and Bncbf17 from Brassica napus. A cold acclimation assay showed that Cbcbf25 was expressed immediately after cold triggering, but this expression was transient, suggesting that it concerns cold acclimation. Our study implies that Cbcbf25 is an analogue of other CBF genes and may participate in cold-response, by for example, controlling the expression of cold-regulated genes or increasing the freezing tolerance of plants.
Shisi Wang,Cancan Xu,Xiaobo Sun,Yifan Zhou,Yaqing Shu,Shangzhou Xia,Zhengqi Lu,Wei Qiu,Xiaofen Zhong,Lisheng Peng 대한신경과학회 2020 Journal of Clinical Neurology Vol.16 No.3
Background and Purpose Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a severe central nervous system disorder mediated by NMDAR antibodies that damages neurons. We investigated the correlation between cytoskeletal autoantibodies and the clinical severity in patients with anti-NMDAR encephalitis. Methods Non-NMDAR autoantibodies were identified by screening matched cerebrospinal fluid (CSF) and the serum samples of 45 consecutive patients with anti-NMDAR encephalitis and 60 healthy individuals against N-methyl-D-aspartate receptor 1-transfected and nontransfected human embryonic kidney 293T cells. Immunocytochemistry was performed to assess antibody binding in rat brain sections and primary cortical neurons. Cell-based assays and Western blotting were applied to identify autoantibodies targeting medium neurofilaments (NFMs). We compared clinical characteristics between patients with NMDAR encephalitis who were positive and negative for anti-NFM-autoantibodies. Results Anti-NFM autoantibodies were detected in both the serum and CSF in one patient (2%) and in the serum only in six patients (13%). No antibodies were detected in the serum of healthy controls (7/45 vs. 0/60, p=0.0016). Four of the seven patients with anti-NFM autoantibodies in serum were children (57%), and three (43%) had abnormalities in brain magnetic resonance imaging. These patients responded well to immunotherapy, and either no significant or only mild disability was observed at the last follow-up. Anti-NMDAR encephalitis did not differ with the presence of anti-NFM autoantibodies. Conclusions Anti-NFM autoantibodies may be present in patients with anti-NMDAR encephalitis, indicating underlying neuronal damage. A large cohort study is warranted to investigate the clinical differences between patients with NMDAR encephalitis according to their anti- NFM antibody status.
Cloning and Functional Analysis of a cDNA Encoding Ginkgo biloba Farnesyl Diphosphate Synthase
Kexuan Tang,Peng Wang,Zhihua Liao,Liang Guo,Wenchao Li,Min Chen,Yan Pi,Yifu Gong,Xiaofen Sun 한국분자세포생물학회 2004 Molecules and cells Vol.18 No.2
Farnesyl diphosphate synthase (FPS; EC2.5.1.1/EC2. 5.1.10) catalyzes the synthesis of farnesyl diphosphate, and provides precursor for biosynthesis of sesquiterpene and isoprenoids containing more than 15 isoprene units in Ginkgo biloba. Here we report the cloning, characterization and functional analysis of a new cDNA encoding FPS from G. biloba. The full-length cDNA (designated GbFPS) had 1731 bp with an open reading frame of 1170 bp encoding a polypeptide of 390 amino acids. The deduced GbFPS was similar to other known FPSs and contained all the conserved regions of trans-prenyl chain-elongating enzymes. Structural modeling showed that GbFPS had the typical structure of FPS, the most prominent feature of which is the arrangement of 13 core helices around a large central cavity. Southern blot analysis revealed a small FPS gene family in G. biloba. Expression analysis indicated that GbFPS expression was high in roots and leaves, and low in stems. Functional complementation of GbFPS in an FPS-deficient strain confirmed that GbFPS mediates farnesyl diphosphate biosynthesis.
Mutations of ARX and non-syndromic intellectual disability in Chinese population
Yufei Wu,Huan Zhang,Xiaofen Liu,Zhangyan Shi,Hongling Li,Zhibin Wang,Xiaoyong Jie,Shao-Ping Huang,Fu-Chang Zhang,Junlin Li,Ke-Jin Zhang,Xiao-Cai Gao 한국유전학회 2019 Genes & Genomics Vol.41 No.1
Mutations of Aristaless-related homeobox (ARX) gene were looked as the third cause of non-syndromic intellectual disability (NSID), while the boundary between true disease-causing mutations and non-disease-causing variants within this gene remains elusive. To investigate the relationship between ARX mutations and NSID, a panel comprising six reported causal mutations of the ARX was detected in 369 sporadic NSID patients and 550 random participants in Chinese. Two mutations, c.428_451 dup and p.G286S, may be disease-causing mutations for NSID, while p.Q163R and p.P353L showed a great predictive value in female NSID diagnosis with significant associations (X2 = 19.60, p = 9.54e−6 for p.Q163R; X2 = 25.70, p = 4.00e−07 for p.P353L), carriers of these mutations had an increased risk of NSID of more than fourfold. Detection of this panel also predicted significant associations between genetic variants of the ARX gene and NSID (p = 3.73e−4). The present study emphasized the higher genetic burden of the ARX gene on NSID in the Chinese population, molecular analysis of this gene should be considered for patients presenting NSID of unknown etiology.
Peng YaFeng,Su XinYu,Hu LiWei,Wang Qian,Ouyang RongZhen,Sun AiMin,Guo Chen,Yao XiaoFen,Zhang Yong,Wang LiJia,Zhong YuMin 대한영상의학회 2021 Korean Journal of Radiology Vol.22 No.9
Objective: To investigate the feasibility of cine three-dimensional (3D) balanced steady-state free precession (b-SSFP) imaging combined with a non-local means (NLM) algorithm for image denoising in evaluating cardiac function in children with repaired tetralogy of Fallot (rTOF). Materials and Methods: Thirty-five patients with rTOF (mean age, 12 years; range, 7–18 years) were enrolled to undergo cardiac cine image acquisition, including two-dimensional (2D) b-SSFP, 3D b-SSFP, and 3D b-SSFP combined with NLM. End-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV), and ejection fraction (EF) of the two ventricles were measured and indexed by body surface index. Acquisition time and image quality were recorded and compared among the three imaging sequences. Results: 3D b-SSFP with denoising vs. 2D b-SSFP had high correlation coefficients for EDV, ESV, SV, and EF of the left (0.959– 0.991; p < 0.001) as well as right (0.755–0.965; p < 0.001) ventricular metrics. The image acquisition time ± standard deviation (SD) was 25.1 ± 2.4 seconds for 3D b-SSFP compared with 277.6 ± 0.7 seconds for 2D b-SSFP, indicating a significantly shorter time with the 3D than the 2D sequence (p < 0.001). Image quality score was better with 3D b-SSFP combined with denoising than with 3D b-SSFP (mean ± SD, 3.8 ± 0.6 vs. 3.5 ± 0.6; p = 0.005). Signal-to-noise ratios for blood and myocardium as well as contrast between blood and myocardium were higher for 3D b-SSFP combined with denoising than for 3D b-SSFP (p < 0.05 for all but septal myocardium). Conclusion: The 3D b-SSFP sequence can significantly reduce acquisition time compared to the 2D b-SSFP sequence for cine imaging in the evaluation of ventricular function in children with rTOF, and its quality can be further improved by combining it with an NLM denoising method.
Yunfei Chen,Qian Shen,Yueyue Wang,Tao Wang,Shaoyan Wu,Ling Zhang,Xu Lu,Fangyuan Zhang,Weimin Jiang,Bo Qiu,Erdi Gao,Xiaofen Sun,Kexuan Tang 한국식물생명공학회 2013 Plant biotechnology reports Vol.7 No.3
Artemisinin is an endoperoxide sesquiterpenelactone isolated from the aerial parts of Artemisia annua L.,and is presently the most potent anti-malarial drug. Owingto the low yield of artemisinin from A. annua as well as thewidespread application of artemisinin-based combinationtherapy recommended by the World Health Organization,the global demand for artemisinin is substantially increasingand is therefore rendering artemisinin in short supply. An economical way to increase artemisinin production is toincrease the content of artemisinin in A. annua. In thisstudy, three key genes in the artemisinin biosynthesispathway, encoding farnesyl diphosphate synthase, amorpha-4, 11-diene C-12 oxidase and its redox partner cytochromeP450 reductase, were over-expressed in A. annuathrough Agrobacterium-mediated transformation. Thetransgenic lines were confirmed by Southern blotting andthe over-expressions of the genes were demonstrated byreal-time PCR assays. The HPLC analysis showed that theartemisinin contents in transgenic lines were increasedsignificantly, with the highest one found to be 3.6-foldhigher (2.9 mg/g FW) than that of the control. Theseresults demonstrate that multigene engineering is aneffective way to enhance artemisinin content in A. annua.
Jun Feng Su,Ruijie Zhang,Xiaofen Hu,Amjad Ali,Zhao Wang 한국화학공학회 2022 Korean Journal of Chemical Engineering Vol.39 No.3
Calcium precipitation induced by Acinetobacter sp. H12 can simultaneously remove F− and Ca2+. Adsorption kinetics and isotherm studies showed that the defluoridation effect improved with the increase of temperature and time. The equilibrium adsorption capacity of calcium precipitation was 7.43 mg g−1 at the initial F− concentration of 10 mg L−1. Moreover, as the pH increased, the F− adsorption capacity decreased and the Ca2+ removal rate gradually increased. The adsorption process was highly fitted to the pseudo-second-order and the Freundlich isotherm. Adsorption thermodynamics analysis demonstrated that the adsorption process was a spontaneous endothermic reaction. The activation adsorption energy was 2.81 kJ mol−1 and the frequency coefficient was 10.88 h−1. EEM, SEM, XPS. XRD results indicated that the defluoridation mechanism may be due to adsorption and co-precipitation. The mechanism of Ca2+ removal may be co-precipitation with F−. It is worth-mentioning that calcium precipitation overcame the problem of adding carbon sources and avoided the microbial safety risk in the effluent. Therefore, a new insight was proposed for simultaneous remediation of Ca2+ and F− from groundwater combining the biological and chemical method.