1.Cloning and prokaryotic expression of a glutathione peroxidase from tick Hemaphysalis longicornis
Chaoying ZHANG ; Guowei GUO ; Zhengchao WANG ; Xiaohong HUANG
Chinese Journal of Zoonoses 2012;(9):861-868
Ticks transmit various diseases to livestock and human beings. The researches on ticks are of great importance in medical and veterinary sciences. To express a glutathione peroxidase gene (HlGPx) from Hemaphysalis longicornis in Escherichia coli (E. coli) so as to provide basis for further studies, the gene was amplified from cDNAs of H. longicornis adult ticks by PCR prompted by information from EST library. The TGA codon encoding selenocysteine in the gene was mutated into the universal genetic code for cysteine,TGC, by site-directed mutagenesis. E. coli was transformed with the recombinant expression vector pGEX-4T-1/HlGPx'(the mutated HlGPx) and induced to express recombinant HlGPx', which was then analyzed by sodium dodecylsulphate-polyacrylamide gel electrophoresis and Western blotting. The result showed that the complete coding sequence of HlGPx was obtained successfully, the deduced polypeptide was 223 aa, with a calculated molecular mass of 25.0 kDa; the recombinant fusion protein was approximately 51 kDa, correspondent to the calculated. The antibody against glutathione S-transferase (GST) recognized the recombinant protein fused with GST in a Western blotting assay, which confirmed the result above-mentioned. In conclusion, the mutated HlGPx was expressed in E. coli successfully and it could be used for further preparation of immune serum and functional analysis.
2.The role of LXRs in the pathogenesis of Alzheimer's disease: a review
Heqiong LIU ; Chaoying MA ; Huicai GUO ; Lei WANG ; Yi LIU
Journal of Preventive Medicine 2023;35(3):215-217
Abstract
Alzheimer's disease (AD) is a neurodegenerative disease characterized by deposition of β-amyloid (Aβ). Liver X receptors (LXRs), a member of the nuclear receptor transcription factor superfamily, are widely expressed in brain, which may be involved in the development and progression of AD. Based on the international and national publications pertaining to the association between LXRs and AD from 2010 to 2022, this review summarizes the advances on the involvement of LXRs in the regulation of cholesterol metabolism, inflammatory response and synapse formation in the pathogenesis of AD was reviewed, so as to provide insights into the prevention and treatment of AD.
3.Expression of focal adhension kinase in circulation endothelial cells from patients with hemorrhagic fever with renal syndrome
Jingyao ZHANG ; Xiuhua ZHAO ; Bo YANG ; Chaoying GUO ; Zhongtao GAI ; Shijun CHEN
Chinese Journal of Infectious Diseases 2010;28(1):30-32
Objective To study the expression of phosphorylated serine 910 of focal adhesion kinase(FAKps910)in circulation endothelial cells(CEC)from patients with hemorrhagic fever with renal syndrome(HFRS).Methods Fifty HFRS patients were involved in the study and divided into mild(n=17),moderate(n=20)and severe(n=13)groups according to patients' conditions. Twenty healthy volunteers were enrolled as control group. CEC were isolated by Percoll density gradient centrifugation method. The expression of FAKps910 in specific APC-CD31+ CEC was measured by flow cytometry. The data were analyzed by one-way ANOVA. Results The positive rates of FAKps910 in CEC were 59.87%±9.58% in early febrile stage patients and 21.14%±2.53% in the healthy controls, while it was 11.64%±2.17%in the later febrile or hypotension stage patients(F=262.31,P<0.01).It gradually restored to normal afterwards. The positive rates of FAKps910 in CEC were 17.45%±2.64%,13.84%±2.54% and 7.47%±2.57%,respectively in mild, morderate and severe groups in the later febrile or hypotension stage, which were all significantly different from that in control group(F=52.642,P<0.01).Conclusions FAK in CEC iS related with the clinical severity of HFRS patients. FAK may be involved in intergrin β3-mediated endothelial injury during Hantavirus infection.
4.Detection of Brain Neurotransmitters in Patients with Vasovagal Syncope with Encephalofluctuogram Technology
Jun XIAO ; Lin WANG ; Chaoying ZHU ; Hongmei LI ; Lun SHU ; Hongling XU ; Yusong GUO ; Chao HU ; Junxian XUE ; Xin CHEN
Chinese Journal of Rehabilitation Theory and Practice 2017;23(3):349-351
Objective To observe the changes of brain neurotransmitters in patients with vasovagal syncope (VVS) with encephalofluc-tuogram technology (ET). Methods From August, 2015 to December, 2016, 30 patients with VVS were selected as case group, 30 controls matched with sex and age were selected from the outpatients without syncope. They were detected the function of gamma aminobutyric acid (GABA), glutamate (Glu), 5-hydroxytryptamine (5-HT), acetylcholine (Ach), norepinephrine (NE) and dopamine (DA) with ET. Results There was no significant difference between the two groups in the values of GABA, Glu, 5-HT, Ach and DA (t<1.680, P>0.05), while the values of NE was higher in the case group than in the control group (t=-3.552, P<0.001). Conclusion VVS may be related to the high level of activity of NE in the brain.
5.Study of the adsorption behaviors of plasma proteins on the single-walled carbon nanotubes nonwoven.
Jie MENG ; Li SONG ; Jie MENG ; Hua KONG ; Chaoying WANG ; Xiaotian GUO ; Haiyan XU ; Sishen XIE
Journal of Biomedical Engineering 2007;24(1):55-60
Single walled carbon nanotubes (SWNT) have attracted increasing research interests for the purpose of biomedical application because they provide not only nanostructured topography, but also chemical composition of pure carbon atoms, as well as ultra high strength and excellent flexibility. Regarding the interactions of nanomaterials to biological systems, non-specific adsorption of plasma proteins is one of the most important issues to be concerned, which plays a crucial role that would determine how biological systems response to the biomaterials. Motivated by application of SWNT materials in biomedical fields, in this study, the adsorption behaviors of plasma proteins on the surface of SWNT nonwoven, prepared directly by floating chemical vapor observation and energy deposition method were investigated by means of scanning electron microscope (SEM), dispersive X-ray (EDX) analysis and ELISA. Results indicated the SWNT non-woven showed a clear adsorption preference of fibrinogen over albumin. There was no human serum albumin detected using above analysis methods on the SWNT nonwoven even incubated in the albumin solution of 4 mg/ml. While more than 0.15 microg of human fibrinogen was detected by ELISA on the SWNT nonwoven with area of 40 mm x 40 mm incubated in the fibrinogen solution of 5 microg/ml. In addition, IgG of sheep-anti-human serum fibrinogen exhibited strong nonspecific adsorption on the surface of SWNT nonwoven. The adsorption behaviors are different significantly from those of other carbon materials and conventional biomaterials. The unique interaction of SWNT nonwoven to plasma proteins is of significance to further studies of blood cells responses.
Adsorption
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Biocompatible Materials
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chemistry
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Blood Proteins
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chemistry
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Nanotubes, Carbon
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chemistry
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Surface Properties