1.The Synchronous Detection and Significance of β-Glucuronidase Both in Serum and Tumorous Tissue in Non-Hodgkin Malignant Lymphoma Patient
Hong ZHANG ; Rui ZHENG ; Bo YANG ; Jing HU ; Bin XU ; Shutao HE ; Anmin YU ; Yingjie LIU
Journal of China Medical University 2001;30(1):4-5,30
Objective: Our purpose was to explore the change regularity of β-glucuronidase (β-G) in body of patients with Non-Hodgkin malignant lymphoma. Methods: β-G was examined synchronously both in the serum and in the tumor tissue of 13 cases patient with Non-Hodgkin malignant lymphoma by using the method of enzymlinked immunsorbent assay (ELISA) and immunohistochemistry separately. Among them, 3 cases were studied by using the immuno electron microscopic technique. Results: β-G was highly expressed both in the serum and tumorous tissue in patients with non-Hodgkin malignant lymphoma and there was obviously difference as compared with the control group (P<0.01). Conclusion: The combined detection with functional and morphological methods to β-G, it may be assistant target to early discovery and early diagnosis of Non-Hodskin malignant lymphoma.
2.Progress and challenges in optical cochlear implant.
Kaiyin ZHANG ; He GUO ; Shan WU ; Yanning WU ; Shutao ZHAO ; Qiuling WANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2016;51(1):73-77
Optical cochlear implant has been occuring as a new cochlear implant which utilizes laser pulses to stimulate hearing. Compared to electronic cochlear implant, it has demonstrated higher spatial selectivity and less radiation scattering, which could lead to higher fidelity cochlear prostheses. At present, most investigations have focused on experiments in vivo. Although a lot of exciting results have been obtained, the mechanisms of laser stimulation is still open. In this paper, a brief review on the recent new findings of optical cochlear implant is given, and possible mechanisms are discussed. In the end, new experimental proposals are suggested which could help to explore the mechanisms of laser-cochlea stimulation.
Cochlear Implantation
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Cochlear Implants
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Hearing Loss
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rehabilitation
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Humans
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Lasers
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Optics and Photonics
3.Expression, purification, stability and transduction efficiency of GST-SOD1-R9 fusion protein.
Jianru PAN ; Lunqiao WU ; Huocong HE ; Lijuan CHEN ; Ying SU ; Lingling LI ; Shutao LIU
Chinese Journal of Biotechnology 2017;33(5):828-837
The fusion of cell permeable peptide TAT and bifunctional antioxidant enzymes, GST (Glutathione sulfur transferase)-TAT-SOD1 (Cu, Zn superoxide dismutase), is an intracellular superoxide scavenger. Compared with SOD1-TAT, GST-TAT-SOD1 has better protective effect on oxidative damage but less transduction efficiency. A novel cell permeable bifunctional antioxidant enzymes with the fusion of GST, SOD1 and polyarginine R9 was constructed for higher transduction efficiency. The full nucleotide sequence of SOD1-R9 was synthesized and inserted into the prokaryotic expression vector pGEX-4T-1 with the GST tag. After the successful construction of the prokaryotic expression vectors of GST-SOD1-R9, the recombinant vector was then transformed into Escherichia coli BL21 (DE3) and the GST-SOD1-R9 fusion protein was produced with the induction of IPTG. The soluble expression of GST-SOD1-R9 fusion protein was combining with the induction temperature and time. The best soluble expression was obtained with the induction temperature of 25 ℃ and the induction time of 11 h. The fusion protein was purified through the combination of 80% ammonium sulfate precipitation and affinity chromatography using glutathione agarose, and verified by SDS-PAGE and special enzymatic activity. The thermal and pH stability of GST-SOD1-R9 fusion protein were analyzed and the SOD and GST activity of fusion protein were proved to be well maintained under physiological conditions. Finally, the transduction efficiency of GST-SOD1-R9 fusion protein was proved to be better than GST-TAT-SOD1 fusion protein (P<0.05). These works establish a foundation for further study of the protective effect of GST-SOD1-R9 fusion protein against oxidative damage.
4.Expression, purification, stability and transduction efficiency of full-length SOD2 recombinant proteins.
Jianru PAN ; Lijuan CHEN ; Huocong HE ; Ying SU ; Xiangling WANG ; Xian LI ; Cuihuang CHEN ; Lunqiao WU ; Shutao LIU
Chinese Journal of Biotechnology 2017;33(7):1168-1177
Superoxide dismutase (SOD) family is necessary to protect cells from the toxicity of reactive oxygen species produced during normal metabolism. Among SODs, manganese-containing superoxide dismutase (Mn-SOD, SOD2) is the most important one. The DNA fragment containing the full nucleotide of full-length human SOD2 was synthesized and inserted into the prokaryotic expression vector pGEX-4T-1 with tag GST. DNA construct was then transformed into Escherichia coli BL21 (DE3) and expression was induced with IPTG at 25 ℃. The recombinant fusion protein GST-SOD2 (46 kDa) was purified from the bacterial lysate by GST resin column affinity chromatography. GST tag was cleaved with thrombin, and a crude SOD2 recombinant protein (25 kDa) was obtained and further purified by heparin affinity chromatography. Activities of the two SOD2 proteins were 1 788 and 2 000 U/mg, respectively. Both SOD2 proteins were stable under physiological condition and cell-penetrating (P<0.05). Our findings open the possibility to study the structure and effects of two full-length recombinant SOD2 proteins.
5.Purification and characterization of two PR-10 protein isoforms from the crude drug of Angelica sinensis.
Xiangling WANG ; Xian LI ; Huocong HE ; Lingling LI ; Di LÜ ; Cuihuang CHEN ; Xiaoqiang YE ; Shutao LIU ; Jianru PAN
Chinese Journal of Biotechnology 2019;35(1):159-168
Two proteins of similar molecular weight (named as ASPR-C-1 and ASPR-C-2) from the crude drug of Angelica sinensis were purified and characterized by 80% ammonium sulfate precipitation, Sephadex G-50 gel filtration chromatography, and DEAE-Sepharose anion exchange chromatography. The molecular weight of ASPR-C-1 and ASPR-C-2 on SDS-PAGE was 17.33 kDa and 17.18 kDa, respectively. They were mainly monomeric in solution, but partially formed dimers and they were glycoproteins with glycosyl content of 2.6% and 8.2%, respectively. Both ASPR-C-1 and ASPR-C-2 were identified to be members of pathogenesis-related 10 family of proteins by matrix-assisted laser desorption ionization time-of-flight mass spectrometry and have ribonuclease activities with the specific activity of 73.60 U/mg and 146.76 U/mg, respectively. The optimum pH of the two isoforms was similar, at about 5.6, while their optimum temperatures were different. The optimum temperature of ASPR-C-1 was 50 ℃, and that of ASPR-C-2 was 60 ℃. Both isoforms presented highest thermal stability at 60 ℃. However, ASPR-C-2 was more thermotolerant than ASPR-C-1. The latter was rapidly inactivated and retained only about 20% residual activity while the former still maintained about 80% of its original activity at a higher treatment temperature (80 to 100 ℃). In addition, Fe²⁺ had an activating effect on the ribonuclease activities of two isoforms while Ca²⁺, Mg²⁺, Zn²⁺, Mn²⁺, Ag⁺, Cu²⁺, EDTA (Elhylene diamine tetraacetic acid), dithiothreitol and sodium dodecylsulphate showed different degrees of inhibition of the enzyme activities. Our findings provide a foundation for further research on the biological function of PR-10 protein from Angelica sinensis.
Angelica sinensis
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Chromatography, Gel
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Chromatography, Ion Exchange
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Electrophoresis, Polyacrylamide Gel
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Enzyme Stability
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Hydrogen-Ion Concentration
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Kinetics
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Molecular Weight
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Protein Isoforms
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Temperature