1.Cloning, expression and protein structure prediction of a novel SCF-TPO fusion gene
Nan LIU ; Yongzhi XI ; Siqi GUO ; Yuying SUN ; Zhihong YUAN ; Jianwu CUI ; Caixia XI ; Fei LIANG ; Fanhua KONG
Chinese Journal of Pathophysiology 2000;0(10):-
AIM: To obtain the high expression of recombinant human stem cell factor - thrombopoitin (SCF-TPO) fusion gene and predict its structure property. METHODS: Tour primers were designed according to known sequence of TPO and SCF to amplify the functional amino acid domain of TPO and SCF by RT- PCR, respectively from fetus hepatocytes. The expression plasmid pET32a/SCF- TPO was constructed by VOE gene fusion technique and expressed in BL21(DE3)plysS. The fusion protein property, such as second structure, flexibility, and hydrophilicity were predicted by DS Gene and Protscale software. RESULTS: The expression vector, pET32a/SCF - TPO was constructed and the high expression of the SCF/TPO fusion protein was obtained, with the expression amount of up to 40% of the total cellular protein. DS Genel .5 and Protscale predict no new antigenicity in fusion protein, and the second structure and ioelectric point have no changes except four amino acids change in first structure. There are high flexibility and low hydrophilieity in the linker peptide. CONCLUSION: High expression of SCF- TPO fusion protein has been obtained and protein prediction shows that the fusion protein design is reasonable, which lay foundation for further study of biological fundation of SCF - TPO fusion protein.
2.Treatment of tibiafibular fractures with rectangle-shaped intramedullary nails
Yuesong WU ; Xinwei WANG ; Jianwu CHEN ; Bocheng XU ; Zhenzhong CUI ; Chongyang ZHAO ; Sulin FAN ; Wenxiao WANG ; Changqing CHEN ;
Academic Journal of Second Military Medical University 2000;0(10):-
Objective:To summarize the 10 year clinical experience of treating tibiafibular fractures with rectangle shaped intramedullary nails(RIN). Methods:From January 1987 to December 1996, 4 682 cases (3 278 male and 1 404 female) of tibiafibular fractures from 9 hospitals were treated with RIN . Three kinds of reduction methods including open reduction, semi open reduction and closed reduction were used during operation. Results:Results showed 2 173 cases (62.89%) got excellent result, 947 got good (27.40%), 214 got moderate (6.19%), 121 got poor (3.50%). The total healing rate was 90.29%. Conclusion:RIN has excellent biological characteristics which can provide a flexible interfixation when treating tibiafibular fractures, and the operation is simple, it also can be used for severe open fractures. RIN is one of the good techniques in treating tibiafibular fractures.
3.Simulated Microgravity can Promote the Apoptosis and Change Inflammatory State of Kupffer Cells
Ge JUN ; Liu FEI ; Nie HONGYUN ; Yue YUAN ; Liu KAIGE ; Lin HAIGUAN ; Li HAO ; Zhang TAO ; Yan HONGFENG ; Xu BINGXIN ; Sun HONGWEI ; Yang JIANWU ; Si SHAOYAN ; Zhou JINLIAN ; Cui YAN
Biomedical and Environmental Sciences 2024;37(10):1117-1127
Objective In this study,we analyzed the transcriptome sequences of Kupffer cells exposed to simulated microgravity for 3 d and conducted biological experiments to determine how microgravity initiates apoptosis in Kupffer cells. Methods Rotary cell culture system was used to construct a simulated microgravity model.GO and KEGG analyses were conducted using the DAVID database.GSEA was performed using the R language.The STRING database was used to conduct PPI analysis.qPCR was used to measure the IL1B,TNFA,CASP3,CASP9,and BCL2L11 mRNA expressions.Western Blotting was performed to detect the level of proteins CASP3 and CASP 9.Flow cytometry was used to detect apoptosis and mitochondrial membrane cells.Transmission electron microscopy was used to detect changes in the ultrastructure of Kupffer cells. Results Transcriptome Sequencing indicated that simulated microgravity affected apoptosis and the inflammatory state of Kupffer cells.Simulated microgravity improved the CASP3,CASP9,and BCL2L11 expressions in Kupffer cells.Annexin-V/PI and JC-1 assays showed that simulated microgravity promoted apoptosis in Kupffer cells.Simulated microgravity causes M1 polarization in Kupffer cells. Conclusion Our study found that simulated microgravity facilitated the apoptosis of Kupffer cells through the mitochondrial pathway and activated Kupffer cells into M1 polarization,which can secrete TNFA to promote apoptosis.