Abstract:Epidermal growth factor (EGF) plays an important role in improving the inflammatory response. In this study, cholera toxin A2 subunit, cell-penetrating peptide trans-transcription activator (TAT) and EGF were fused and recombinant protein EGF-CTA2-TAT were expressed by Escherichia coli. Two methods of renaturation, affinity chromatography and dialysis, were selected to renature the protein. The renaturation efficiency of these two methods were compared. Activities of recombinant protein on promoting BALB/c 3T3 cells proliferation in vitro were also studied. After that, mice model of neuroinflammation was established by using the Lipopolysaccharide (LPS) intraperitoneal injection. The purified and renatured recombinant EGF-like protein was administered intranasally for neuroinflammation treatment. The Morris water maze (MWM) test was used to evaluate the therapeutic effects. Our results showed that recombinant EGF-like protein had a therapeutic effect on LPS-induced neuroinflammation in mice, and could remarkably improve the learning and memory ability of mice. The recombinant EGF-like protein renatured and purified by affinity chromatography in one step shows the advantages of simple process, high yield, excellent activity and low cost, providing a reference for mass production of such active proteins.

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	1. Study on the renaturation process of epidermal growth factor-like protein and its therapeutic effect on neuroinflammation
	Yang Jingru,He Huahong,He Huafeng,Wang Huaqian
	Biology 25 March 2023
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	Abstract:Epidermal growth factor (EGF) plays an important role in improving the inflammatory response. In this study, cholera toxin A2 subunit, cell-penetrating peptide trans-transcription activator (TAT) and EGF were fused and recombinant protein EGF-CTA2-TAT were expressed by Escherichia coli. Two methods of renaturation, affinity chromatography and dialysis, were selected to renature the protein. The renaturation efficiency of these two methods were compared. Activities of recombinant protein on promoting BALB/c 3T3 cells proliferation in vitro were also studied. After that, mice model of neuroinflammation was established by using the Lipopolysaccharide (LPS) intraperitoneal injection. The purified and renatured recombinant EGF-like protein was administered intranasally for neuroinflammation treatment. The Morris water maze (MWM) test was used to evaluate the therapeutic effects. Our results showed that recombinant EGF-like protein had a therapeutic effect on LPS-induced neuroinflammation in mice, and could remarkably improve the learning and memory ability of mice. The recombinant EGF-like protein renatured and purified by affinity chromatography in one step shows the advantages of simple process, high yield, excellent activity and low cost, providing a reference for mass production of such active proteins.
	TO cite this article:Yang Jingru,He Huahong,He Huafeng, et al. Study on the renaturation process of epidermal growth factor-like protein and its therapeutic effect on neuroinflammation[OL].[25 March 2023] http://en.paper.edu.cn/en_releasepaper/content/4759631


	2. A review: Inhibitors and activators for paraoxonase 1
	Li Yan,Tang Meiling,Chen Gang,Mu Xiaojing,Huang Haiming
	Biology 23 April 2020
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	Abstract:Paraoxonase 1 (PON1) is a calcium-dependent hydrolase with many physiological functions. The decrease of PON1 activity is related to many diseases. The research about PON1 is a hot topic, and its activators and inhibitors are expected to help in our diets and in choose of drugs. The papers about PON1 activators and inhibitors were collected and classified. The PON1 activators and inhibitors are grouped into four categories: in-vivo activators, in-vitro activators, in-vivo inhibitors and in-vitro inhibitors. For each activator or inhibitor, the activating effect or inhibiting effect along with the methods used was reviewed. Up to now, we found from literatures that there were 16 samples demonstrating activating effects on PON1 in-vivo and 5 compounds showed in-vitro activation. Of the 16 in-vivo activators there were 4 plant extracts and 12 pure components. Seven in-vivo inhibitors and 71 in-vitro inhibitors have been reported in publications. Additionally, smoking, drinking and treatment with exogenous gonadotropins induced decrease of PON1 activity. The summary of the activators and inhibitors are beneficial to reveal the structure-activity relationship, and also it provides convenience to look up the activating or inhibiting effects of the compounds on PON1 at a glance.
	TO cite this article:Li Yan,Tang Meiling,Chen Gang, et al. A review: Inhibitors and activators for paraoxonase 1[OL].[23 April 2020] http://en.paper.edu.cn/en_releasepaper/content/4751765

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	3. The research in equilibrium study and diversified models of thifensulfuron methyl in several common pure solvents and mixture organic solvents from (278.15 to 333.15) K
	Yuhao Zhang,Yemin Yu,Yonghong Hu,Renlun Deng,Wenge Yang,Tao Li,Yang Cao
	Biology 10 May 2017
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	Abstract:IThifensulfuron methyl is a kind of herbicide and an important chemical intermediate product, Data on corresponding solid?liquid equilibrium of thifensulfuron methyl in different solvents is essential for a preliminary study of industrial applications. In this study, the solubility of thifensulfuron methyl in water, methanol, ethanol, butyl alcohol, isopropanol, acetone, acetonitrile, hexane and ethyl acetate pure solvents and the solubility in (ethanol + ethyl acetate) mixture solvents was measured in the temperature range from 278.15 K to 328.15 K. We use the gravimetric method get the solubility data, contact with the modified apelblat equation and λh equation. We can calculate and analysis thermodynamic parameters (including enthalpy, entropy, and gibbs energy) by the van't hoff analysis and gibbs equation. Calculations proves that the modified apelblat model has advantages than the other two models. In addition, the calculated thermodynamic parameters indicated that in each studied solvent the dissolution of thermodynamic models is endothermic, nonspontaneous, and an entropy-driven process.
	TO cite this article:Yuhao Zhang,Yemin Yu,Yonghong Hu, et al. The research in equilibrium study and diversified models of thifensulfuron methyl in several common pure solvents and mixture organic solvents from (278.15 to 333.15) K[OL].[10 May 2017] http://en.paper.edu.cn/en_releasepaper/content/4732331


	4. Identification of a Flavonoid Glucosyltransferase Involved in 7-OH Site Glycosylation in Tea plants
	Dai Xinglong,Zhuang Juhua,Wu Yingling,Wang Peiqiang,Zhao Guifu,Liu Yajun,Jiang Xiaolan,Gao Liping,Xia Tao
	Biology 22 April 2017
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	Abstract:Flavonol glycosides, which are often converted from aglycones catalyzed by UDP-glycosyltransferases (UGTs), play important roles for the health of plants and animals. In present study, CsUGT75L12, a gene encoding a flavonoid 7-O-glycosyltransferase, was identified in tea plants. Recombinant CsUGT75L12 protein displayed glycosyltransferase activity with multiple phenolic compounds including flavanone, flavone, flavonol and isoflavone on the 7-OH position. In relative comparison to wild-type seeds, the content of flavonol-glucosides levels increased in seeds of Arabidopsis overexpressing CsUGT75L12. In order to determine the key amino acid residues responsible for the catalytic activity of the protein, a series of site-directed mutagenesises and enzymatic assays were performed based on the 3D structural modeling and docking analysis. These results suggested that residue Q54 was predicted to be a double binding site functioning as a sugar receptor and donor. Residues H56 and T151 corresponding to the basic active residues H20 and D119 of VvGT1, were not irreplaceable for CsUGT75L12. In addition, residues Y182, S223, P238, T239, and F240 were demonstrated to be responsible for a "reversed" sugar receptor binding model. The results of triple substitution confirmed that the function of residues P238, T239 and F240 may substitute or compensate with each other for the flavonoid 7-O-glycosyltransferase active.?????
	TO cite this article:Dai Xinglong,Zhuang Juhua,Wu Yingling, et al. Identification of a Flavonoid Glucosyltransferase Involved in 7-OH Site Glycosylation in Tea plants[OL].[22 April 2017] http://en.paper.edu.cn/en_releasepaper/content/4728169


	5. The effect of centrally administered apelin-13 on interdigestive gastric motility in conscious rats
	LV Shuangyu,QIN Yaojun,WANG Ningbo,YANG Yanjie,CHEN Qiang
	Biology 02 August 2012
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	Abstract:Apelin, a novel neoropeptide, was identified as the endogenous ligand for APJ receptor. The present study was designed to explore the effect apelin-13 on interdigestive gastric motility in conscious rats. Apelin-13 was synthesized by the solid-phase peptide synthesis method. After recovery from the operation of intracerebroventricular (i.c.v.) cannula implantation, the strain gauge transducers were implanted on the serosal surface of the antrum in Wistar rats. The migrating motor complex (MMC) was instantly recorded by the Power-Lab recording system. The results show that, comparing with control group (5.17±0.93), the frequency of the phase Ⅲ-like activity (c/h) of the 10 nmol apelin-13 group (7.67±1.33) and 30 nmol apelin-13 group (8.58±1.66) were both increased, showing a statistically significant difference (P < 0.01, P < 0.01, respectively). However, frequency of the phase Ⅲ-like activity of 3 nmol apelin-13 group (6.83±0.98) had no significant difference (P > 0.05). Comparing with control group (97.41±16.88), the %motor index (MI) of the 10 nmol apelin-13 group (143.93±26.27) and 30 nmol apelin-13 group (155.09±24.02) were both increased, indicating a statistically obvious difference (P < 0.01, P < 0.01, respectively). However, the %MI of 3 nmol apelin-13 group (120.14±3.00) showed no significant difference (P > 0.05). Our study first indicates that i.c.v. injection of apelin-13 increased the interdigestive antrum MMC in conscious rats, which has an important physiological significance for further study the modulatory effect of apelin-13 on the digestive tract motility.
	TO cite this article:LV Shuangyu,QIN Yaojun,WANG Ningbo, et al. The effect of centrally administered apelin-13 on interdigestive gastric motility in conscious rats[OL].[ 2 August 2012] http://en.paper.edu.cn/en_releasepaper/content/4485939


	6. Effects of Wingless / Wnt1 on the Development of Tribolium castaneum
	ZhuoyuLi,YananPeng,BinLi,ChengjunLi
	Biology 25 June 2012
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	Abstract:Known as highly conserved during evolution, the Wnt signaling pathway plays a vital role in regulating animal embryonic axis, embryonic differentiation, and deciding cell polarity and maintaining adult dynamic equilibrium. Mutations or deregulations of its components can cause the occurrence of carcinoma. We studied the role of Wingless/Wnt1 during larva-adult development of the red flour beetle, Tribolium castaneum, with dsRNA-mediated Wingless(Wg)/Wnt1 gene being knocked down. The treated late larvae metamorphosed into pupae with drastically increased wing interval and decreased wing width (P<0.01). The pursuant pupa-adult eclosion was also severely affected and most of pupae died during this period. The qPCR result showed that the mRNA level of Cadherin-like and Smoothened (Smo) genes were up-regulated greatly, and that of armadillo-2 was slightly higher , after Wingless/Wnt1 gene was knocked down. We drew the conclusion that Wnt-1 signaling pathway is closely related to the proper wing development and adult metamorphosis of Tribolium. In addition, the elevated expression of Cadherin-like and Armadillo-2 may be accountable for the reduced wing width and enlarged wing interval caused by wg gene silencing, because those alterations can either enhance cell adhesion or change cell morphology. Importantly, the up-regulation of smo gene indicates that Hedgehog signaling pathway may be affected by the RNAi of wg and involved in the abnormal or lethal phenotypes observed in our experiment.??
	TO cite this article:ZhuoyuLi,YananPeng,BinLi, et al. Effects of Wingless / Wnt1 on the Development of Tribolium castaneum[OL].[25 June 2012] http://en.paper.edu.cn/en_releasepaper/content/4482937


	7. Purification，Spectroscopic and Thermodynamic Characterization of Metallo-β-Lactamase L1 from Stenotrophomonas maltophilia
	Hui-Zhou GAO,Xiao-Yan YAN,Qi YANG,Xia YANG,Ji-Li FENG,Sheng-Li GAO,Ke-Wu YANG
	Biology 18 September 2010
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	Abstract:In an effort to probe the antibiotic resistance in bacteria, metallo-β-lactamase L1 from stenotrophomonas maltophilia was overexpressed in Escherichia coli, purified by ion-exchange chromatography and characterized by mass spectroscopy, fluorescence and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The matrix-assisted laser desorption ionization and time-of-flight (MALDI-TOF) mass spectroscopy result shows that a monomeric molecular mass of L1 is 28.8 kDa. This paper first report the determination of the thermodynamic enthalpy change of L1-catalyzed hydrolysis reaction of penicillin G and the enthalpy change value in average at 298.15K is 76.8 kJ/mol.
	TO cite this article:Hui-Zhou GAO,Xiao-Yan YAN,Qi YANG, et al. Purification，Spectroscopic and Thermodynamic Characterization of Metallo-β-Lactamase L1 from Stenotrophomonas maltophilia[OL].[18 September 2010] http://en.paper.edu.cn/en_releasepaper/content/4386049


	8. Expression, Purification and Characterization of Malate dehydrogenase from Thermus thermophilus HB27
	Liu Jun,Lv Zhengbing,Meng Fanguo,Zhang Yaozhou
	Biology 16 November 2009
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	Abstract:Malate dehydrogenase (MDH, EC 1.1.1.37) is an essential metabolic enzyme in tricarboxylic acid cycle. The gene of the malate dehydrogenase of Thermus thermophilus HB27 was cloned and expressed in E.coli. The 984 bp gene encoded a homodimer enzyme of 35.39 kDa per subunit. The enzyme was purified by heat purification and metal chelate affinity chromatography. The relative activity was measured in different grades of pH value and temperature. The results showed that the optimal pH value was approximately 11.5 and the optimal temperature was 80℃. Our results may provide useful information regarding the biochemical properties of the MDHs from thermophilic bacteria.
	TO cite this article:Liu Jun,Lv Zhengbing,Meng Fanguo, et al. Expression, Purification and Characterization of Malate dehydrogenase from Thermus thermophilus HB27[OL].[16 November 2009] http://en.paper.edu.cn/en_releasepaper/content/36725


	9. Novel fluorescent proteins generated by de novo synthesis method and their characterization
	Sun Tingting,Yi Ke,Zhu Cong,Wang Jinchun,Gao Xiaolian,Guo Jiangfeng
	Biology 28 September 2009
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	Abstract:Up to now the method to get fluorescent proteins (FPs) variants is focused on random mutation and site-directed mutagenesis. Herein we can not forecast and get the special expected mutated site at our will for random mutation. Though site-directed solved this problem, its inherent limitation that it is difficult to get amount mutation at the same time and it is a money-spending and time-consuming method hindered its application. In this article we used a high throughput and relatively cheap technology- microfludic PicoArray method for the simultaneous de novo synthesis fluorescent proteins（FPs）DNA sequences and reassemble sequences successively to get massive variants of fluorescent proteins. The result showed that five FPs are unique and have properties different to the others FPs existed and within these five new FPs, three FPs with different hues are belong to red fluorescent proteins which emission wavelength are longer than 550 nm and the other two locate at green fluorescent protein region. The native gel combined with Mass spectrum showed that all the five FPs are monomer with molecular weight are between 25 kD and 35 kD and they have relative stronger fluorescence intensity comparing with former blue fluorescent proteins, so they will be potential tools in molecular and medical studying fields.
	TO cite this article:Sun Tingting,Yi Ke,Zhu Cong, et al. Novel fluorescent proteins generated by de novo synthesis method and their characterization[OL].[28 September 2009] http://en.paper.edu.cn/en_releasepaper/content/35543


	10. Hydrophobic Variety of Chromatin in Cellular processes
	Xiao-en Wang
	Biology 06 November 2008
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	Abstract:The discrepancies of molecular structure between DNA and RNA result to their different nature. Deoxyribose constituting DNA is more hydrophobic than ribose constituting RNA. On the chemical principle of similar substances can be soluble with each other (SSSEO), here, I propose that the DNA, nucleoplasm, and decondensational state of chromatin are more hydrophobic than the RNA, cytoplasm, and condensational state, respectively. Based on this point, many biological processes, such as the disappearance and reappearance of nuclear envelope during division of eucaryote, the spontaneous exit of RNA to cytoplasm after synthesis within nucleus, and the relation between chemical modifications of chromatin and some important cellular processes, may be well explicated.
	TO cite this article:Xiao-en Wang. Hydrophobic Variety of Chromatin in Cellular processes[OL].[ 6 November 2008] http://en.paper.edu.cn/en_releasepaper/content/25495

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