4.Expression and activities analysis of a fusion protein CREKA/tTF
Yi SU ; Jianghua YAN ; Shengyu WANG ; Jie HE ; Min YE
Chinese Journal of Biochemical Pharmaceutics 2010;31(2):94-97
Purpose To prepare a novel fusion protein of CREKA and tTF as a universal carrier targeting to cancer,and to analyze its activities.Methods CREKA and tTF gene were acquired by PCR,and inserted into plasmid pET22b(+)to construct recombinant plasmid CREKA/tTF/pET22b(+),and the fusion gene was expressed in E.coli BL21.The fusion protein Wag purified through Nickel-affinity chromatography column.After purifying,the fusion protein was refold by subsequent dialysis.The activities of the fusion proteins were measured by coagulation timing and quantitative fluorescence test in vitro.Results The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was obtained.The fusion protein was highly expressed in E.coli BL21.The coagulation of the fusion protein Was determined by the coagulation test.And the capability of the fusion protein effectively binding to clotted plasma proteins is identified in quantitative fluorescence test.Conclusion The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was built.The fusion protein CREKA/tTF with both TF and CREKA activity was successfully obtained.
5.Prostate cancer: diagnostic value of quantitative analysis by dynamic contrast-enhanced MR at 3.0 T
Wei HE ; Yi LIU ; Jianyu LIU ; Min LU
Chinese Journal of Radiology 2014;48(3):215-218
Objective To investigate the diagnostic value of the quantitative analysis parameters of dynamic contrast-enhanced MRI (DCE-MRI) for prostate cancer.Methods MR examination were performed in 26 patients and correlations were made with surgical pathology following radical prostatectomy.According to the localization of pathologic specimens,ROI were drawn on areas of cancerous regions,noncancerous regions in peripheral zone and central gland to measure the values of Ktrans,kep and Ve.The values of the three parameters in different tissue were compared with Kruskal-Wallis H and Mann-Whitney U.Results Twenty six patients had prostate cancer confirmed by pathology.Data from 468 assessable regions of prostate were acquired,including 115 cancerous regions and 142 noncancerous regions in central gland,68 cancerous regions and 143 noncancerous regions in peripheral zone.Prostate cancer showed a multifocal distribution.The K,kep and Ve values were (1.04 ± 0.26)/min,(1.43 ± 0.46)/min,(0.76 ±0.12) respectively for cancerous regions in central gland,and (0.82 ±0.19)/min,(1.12 ±0.26)/min,(0.75 ± 0.14) respectively for noncancerous regions in central gland,(0.95 ±0.31)/min,(1.31 ±0.51)/min,(0.76 ± 0.13) respectively for cancerous regions in peripheral zone and (0.32 ± 0.07)/min,(0.52 ± 0.13)/min,(0.64 ± 0.14) respectively for noncancerous regions in peripheral zone.The differences among the three parameters were statistically significant (x2 =316.008,297.985,63.681,P <0.01).The Ktrans,kep values of cancerous regions were significantly higher than the corresponding values of noncancerous regions,respectively.Conclusion Quantitative analysis parameters of DCE-MRI contribute to the diagnosis of prostate cancer,including cancer located in central gland.
6.Genetic Engineering Progresses in Plant Resistance to Salt Stress
Yi-Min HE ; Hong-Juan NIAN ; Li-Mei CHEN ;
China Biotechnology 2006;0(03):-
Salinity is the main limitation factor for plant growth and crop production.Many approaches to enhance plant resistance to salinity by genetic engineering have been developed.Over-expressions of salt-tolerance related genes encoding proteins involved in signal transduction pathways,ion channels and compatible solutes synthesis for the stabilization of biological structures under salinity stress are the most often used strategies.The recent progresses in genetic engineering to improve salt tolerance in plants and the possible problems in researches was reviewed.
7.Hybrid repair for dissection involving the aortic arch
Yi SI ; Yi HE ; Min TANG ; Hao LIU ; Fangbao DING ; Chunrong BAO ; Ju MEI
Chinese Journal of General Surgery 2016;31(8):624-627
Objective To summarize the experience of hybrid repair performed in high risk patients with dissection involving the aortic arch.Methods From Sep.2007 to Mar.2015,hybrid repair was performed in 33 high risk patients with dissection involving the aortic arch including acute (n =8),subacute (n =15),or chronic (n =10) cases.Descripitive statistics were computed for continuous and categorical variables.Results There were 22 male and 11 female patients with a mean age of(69 ± 10) years,and ASA Physical Status Ⅲ-Ⅳ.Simultaneous (n =27) and staged (n =5,mean interval 5.0 ± 1.3 days)endovascular repair were performed via femoral artery.The technical success rate was 100%.The average hospital stay was (16 ±6) days.One case died of cerebral infraction.There were two with strokes,one with pneumonia and two with renal failure as complications.Median follow-up was 47 months (3-66 months).There were four deaths with two were related to aortic artery.Endoleak was found in 3 during follow-up.One type Ⅰ endoleak was cured after remedy hybrid repair.Conclusions Hybrid repair performed in patients at high risk with dissection involving the aortic arch is less invasive with favorable medium and long-term outcomes.
8.Effect of sulfatide-activated typeⅡnatural killer T cells on airway inflammation in a murine model of asthma
Qing HE ; Qiaoyu YANG ; Min LIU ; Hanxiang NIE ; Guqin ZHANG ; Xuhong DING ; Yi HUANG ; Hongying YU
Chinese Journal of Immunology 2014;(11):1452-1456,1461
Objective:To investigate the effect of type ⅡNKT cells activated by sulfatide on airway inflammation in a murine model of asthma.Methods:Thirty-two BALB/c mice were randomly divided into four groups:normal control group ( n=8 ) , asthma group (n=8),sulfatide treatment group (n=8) and adoptive transfer group (n=8).The murine model of asthma was established by sensitization with intraperitoneal injection of ovalbumin ( OVA) and intranasal challenge in all animals except for the normal control group where PBS was used instead.Intraperitoneal injection of sulfatide in a sulfatide treatment group, adoptive transfer of sulfatide-activated typeⅡNKT cells in adoptive transfer group and PBS in asthma group were carried out 1 hour before the first challenge.PBS was used for intraperitoneal administration in the normal control group.Lung histology and goblet cell hyperplasia were analyzed by HE or PAS staining.Differential cell count in bronchial alveolar lavage ( BALF) was measured by May-Gruenwald Giemsa;levels of OVA-specific IgE in serum and L-4,IL-5 in BALF were measured by ELISA.The percentages of lung type Ⅱ NKT cells,IL-4+and IFN-γ+typeⅡNKT cells were detected by flow cytometry.Results:Inflammatory cell infiltration in lung tissue and goblet cell hyperplasia in the airway were decreased in sulfatide treatment group and adoptive transfer group.Percentages of eosinophil in BALF,level of OVA-specific IgE in serum,and levels of IL-4,IL-5 in BALF in sulfatide treatment group and adoptive transfer group were significantly lower than those in asthma group (all P<0.05).The percentages of lung IL-4+and IFN-γ+typeⅡNKT cells in sulfatide treatment group was significantly higher than those in asthma group ( P<0.01 ).Conclusion: Type Ⅱ NKT cells activated by sulfatide may inhibit airway inflammation in a murine model of asthma.
9.Expression of estrogen and androgen receptors on aneurysm walls and their effects in patients with intracranial aneurysm
Feng YE ; Chao YOU ; Hong XU ; Min HE ; Aiguo LI ; Yi LIU
Chinese Journal of Cerebrovascular Diseases 2017;14(6):308-312
Objective To investigate the significance of androgen and estrogen receptor expression levels on aneurysm walls.Methods From November 2007 to June 2016,32 patients received craniotomy for clipping intracranial aneurysms in the West China Hospital,Sichuan University were enrolled prospectively.Nineteen intracranial aneurysm walls and 26 superficial temporal artery branches were obtained (a total of 45 qualified specimens).Immunohistochemical method was used to detect the superficial temporal artery branches and smooth muscle layer of intracranial aneurysm wall and the expression levels of estrogen receptor-α,β and androgen receptors.Image Pro Plus 6.0 software was used to analyze and detect the integral optical density values of the positive cell expression levels.The χ2 test and rank-sum test were used for statistical analysis.Results The median (M) and interquartile range (P25,P75) of the expression levels of estrogen receptor-α,β of the intracranial aneurysm walls were 3 049 (2 112,5 554) and 4 364 (2 314,5 667) respectively.They were lower than 6 544 (3 507,10 103) and 6 972 (5 694,10 024) of the superficial temporal artery branches.The expression level of androgen receptor of aneurysm wall was 3 299 (1 375,4 895),it was higher than 1 130 (794,1 922) of the superficial temporal artery branches.There was significant difference between the two groups (all P<0.05).Conclusion The decreased expression levels of estrogen receptor-α,β and the increased expression level of androgen receptor in the cerebrovascular walls may promote the progress of intracranial aneurysms,however,the specific mechanism needs further study.
10.The Effect of p65 siRNA on the Expression of Pro-Inflammatory Factors under Conditions Mimicking Ischemia/Reperfusion in Human Umbilical Vein Endothelial Cells
Min ZENG ; Zhiyong WU ; Yin ZHENG ; Yangli HE ; Yi FEI ; Xiaojun LIU
Tianjin Medical Journal 2014;(2):97-100
Objective To investigate the effect and regulation mechanism of mimic ischemia/reperfusion (I/R) cul-ture of human umbilical vein endothelial cells (HUVECs) on levels of tumor necrosis factor(TNF)-αand intercellular adhe-sion molecule (ICAM)-1. Methods HUVECs were randomly divided into four groups:control group (normal media cell cul-ture+control siRNA transfection), mimic I/R+control siRNA transfection group (HUVECs were transfected with control siR-NA, for 48 h ,and then received mimic ischemic media culture for 30 min followed by normal media culture for 4 h), normal culture+p65 siRNA transfection group and mimic I/R+p65 siRNA transfection group. The expression levels of TNF-αand ICAM-1 mRNA and protein were determined by real-time PCR and enzyme linked immunosorbent assay (ELISA), respec-tively. Results The mRNA and supernatant protein levels of TNF-αand ICAM-1 were significantly increased in mimic I/R culture group (4.96±0.16 and 3.33±0.30)μg/L than those of other tree groups (P<0.05). The level of ICAM-1 mRNA was significantly higher in I/R+p65 siRNA transfection group (1.87±0.21)μg/L than that of control group (1.58±0.15) μg/L and normal culture+p65 siRNA transfection group [(1.69±0.21)μg/L, P<0.05]. The levels of TNF-αand ICAM-1proteins were (329.98 ± 12.18) μg/L and (654.74 ± 64.79) μg/L in mimic I/R+control siRNA transfection group, which were significantly higher than those of other three groups (P<0.05). The levels of TNF-αand ICAM-1 proteins were (129.65±22.42)μg/L and (185.76±11.27)μg/L in mimic I/R+p65 siRNA transfection group, which were significantly lower than those of contro group [(183.50±11.77)μg/L and (280.43±13.76)μg/L, P<0.05]. Conclusion The silencing of p65 through transfection of p65 siRNA in HUVECs inhibited mimic I/R-induced mRNA and protein expressions of TNF-αand ICAM-1.