2.Cloning and bioinformatic analysis and expression analysis of beta-glucuronidase in Scutellaria baicalensis.
Shuang-shuang GUO ; Lin CHENG ; Li-min YANG ; Mei HAN
China Journal of Chinese Materia Medica 2015;40(22):4370-4377
The β-Glucuronidase gene (sbGUS) cDNA firstly from Scutellari abaicalensis leaf was cloned by RT-PCR, with GenBank accession number KR364726. The full length cDNA of sbGUS was 1 584 bp with an open reading frame (ORF), encoding an unstable protein with 527 amino acids. The bioinformatic analysis showed that the sbGUS encoding protein had isoelectric point (pI) of 5.55 and a calculated molecular weight about 58.724 8 kDa, with a transmembrane regions and signal peptide, had conserved domains of glycoside hydrolase super family and unintegrated trans-glycosidase catalytic structure. In the secondary structure, the percentage of alpha helix, extended strand, β-extended and random coil were 25.62%, 28.84%, 13.28% and 32.26%, respectively. The homologous analysis indicated the nucleotide sequence 98.93% similarity and the amino acid sequence 98.29% similarity with S. baicalensis (BAA97804.1), in the nine positions were different. The expression level of sGUS was the highest in root based on a real-time PCR analysis, followed by flower and stem, and the lowest was in stem. The results provide a foundation for exploring the molecular function of sbGUS involved in baicalcin biosynthesis based on synthetic biology approach in S. baicalensis plants.
Amino Acid Sequence
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Base Sequence
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Cloning, Molecular
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Computational Biology
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Glucuronidase
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chemistry
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genetics
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metabolism
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Molecular Sequence Data
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Open Reading Frames
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Phylogeny
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Plant Proteins
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chemistry
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genetics
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metabolism
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Protein Structure, Secondary
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Scutellaria baicalensis
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chemistry
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enzymology
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genetics
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Sequence Alignment
3.Effect of hypoxia on adhesion and invasion of human cervical cancer cell HeLa
Shuang LIN ; Li LI ; Ping YI
Journal of Third Military Medical University 2003;0(18):-
Objective To investigate the potential effect of hypoxia on adhersion and invasion of human cervical carcinoma cells HeLa. Methods Reverse transcription-polymerase chain reaction (RT-PCR) was used to investigate the level of HIF-1? mRNA and E-cadherin mRNA of HeLa cells cultured under normoxia, 5%O2, 3%O2, 1%O2. The invasiveness of HeLa cells was investigated by transwell chamber assay. Results Under the condition of normoxia, 5%O2, 3%O2 and 1%O2, the optical density ratio of HIF-1? mRNA/?-actin was (0.423?0.116), (0.764?0.111), (1.687?0.126) and (0.215?0.099), showing that the level of HIF-1? mRNA had a tendency of first inceasing and then decreasing with the decreasing oxygen content. The optical density ratio of E-cadherin/?-actin was (1.045?0.171), (0.719?0.114), (0.386?0.105) and (0.157?0.063), showing a decreasing tendency with the decreasing of oxygen content. The invasiveness of HeLa cells was (112.5?20.7), (254.3?27.5), (333.6?45.2) and (124.5?24.6), showing that the invasiveness of HeLa cells had a tendency of first inceasing and then decreasing with the decreasing oxygen content. Conclusion Moderate hypoxia up-regulates the mRNA levels of HIF-1?, and down-regulates the mRNA levels of E-cadherin, and enhances the tumor invasiveness. The possible mechanism is the increasing mRNA levels of HIF-1?.
4.Potassium channel blocker(4-AP)enhances the radiosensitivity of human cervical cancer cell line HeLa to ~(252)Cf neutron ray
Shuang LIN ; Li LI ; Yuan LIU
Journal of Third Military Medical University 2003;0(10):-
Objective To investigate the potential effect of 4-amino-pynidine(4-AP)on human cervical carcinoma cell line HeLa to 252Cf neutron ray.Methods HeLa cells were treated with 4-AP(0,1,5,10,20 mmol/L,respectively)and then exposed to 0.67 Gy 252Cf neutron ray.The effect of 4-AP on proliferation of HeLa cells was assessed by MTT assay.Cell apoptosis were detected by flow cytometry.Reverse transcription polymerase chain reaction(RT-PCR)and Western blotting were used to investigate the levels of HIF-1? mRNA and protein of HeLa cells treated with 4-AP(0,20 mmol/L)and 0.67 Gy 252Cf neutron ray.Results After HeLa cells treated with 0,1,5,10,20 mmol/L 4-AP were exposed to 0.67 Gy 252Cf neutron ray for 48 h,their inhibitory rate was 0,(38.81 ?3.45)%,(46.63?3.88)%,(63.58?6.19)%,(77.51?8.87)%,and their apoptotic rate was(3.15?0.85)%,(8.01?1.19)%,(16.00?1.58)%,(47.20?3.18)%,(62.56?4.27)%,respectively,suggesting 4-AP may inhibit the proliferation of HeLa cells and induce their apoptosis in a dose-dependent manner(P0.05).However,after treated with 20 mmol/L 4-AP and exposed to 0.67 Gy 252Cf neutron ray for 24,48 and 72 h,the optical density ratio of HIF-1? mRNA/?-actin of HeLa cells was(0.401?0.121),(0.364?0.142),(0.257?0.137),(0.165?0.099),while the optical density ratio of HIF-1? protain/?-actin were(0.879?0.117),(0.645?0.115),(0.312?0.114),(0.130?0.118),showing that the levels of HIF-1? mRNA and protein both declined(P
5.Significance of HIF-1? and E-cadherin in cervical carcinoma
Shuang LIN ; Li LI ; Yun WANG
Journal of Third Military Medical University 2003;0(10):-
Objective To study the significance of the expressions of HIF-1? and E-cadherin in cervical carcinoma. Methods The expressions of HIF-1? and E-cadherin (E-cad) were determined in the clinical specimens of 109 cases of cervical carcinoma, 16 cases of cervical intraepithelial neoplasm and 10 cases of normal cervical tissue with immunohistochemical assay. Results There was nearly no expression of HIF-1? in the normal cervical tissue. The positive expression of HIF-1? and E-cad were 81.4% (70/86) and 28.6% (25/86) respectively in the tissue of invasive cervical carcinoma. The rate of positive expression of HIF-1? was significantly higher in invasive cervical carcinoma than in normal cervical tissue and cervical intraepithelial neoplasm (P
7.Investigation on level of DDT accumulation in women and children of a polluted area.
Shuang LI ; Chao-lin LI ; Yan-rang WANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2010;28(5):353-355
Adult
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Child
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DDT
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analysis
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blood
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Environmental Pollution
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prevention & control
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Female
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Humans
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Male
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Milk, Human
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chemistry
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Young Adult