4.Caspase-8 dependent osteosarcoma cell apoptosis induced by proteasome inhibitor MG132
Xiaobo YAN ; Xiang GAO ; Jie FENG ; Zhongli SHI ; Disheng YANG
Chinese Journal of Pathophysiology 1986;0(02):-
AIM: To investigate the effects of proteasome inhibitor Z-LLL-CHO(MG132) on human osteosarcoma cell line MG-63 and its possibly mechanism.METHODS: After treated with different concentration of MG132,the morphological change,ultrastructral morphology,cell viability,cell apoptosis,gene transcription and protein expression in MG-63 cells were accessed by fluorescence microscope,electron microscope,MTT assay,agrose gel electrophoresis,FCM,RT-PCR and Western blotting.RESULTS: Proteasome inhibitor MG132 was an effective inducer of apoptosis in human osteosarcoma MG-63 cells.Not only apoptotic changes,but also cell arrest at G2-M-phase,the accumulation of p27kip1,the accumulation of activated caspase-8 and increased ratio of Bax∶Bcl-2 were observed.However,to normal human diploid fibroblast cells,MG132 did not show apoptosis-inducing effect.CONCLUSION: Apoptosis induced by MG132 may be caspase-8,p27kip1and bcl-2-related.
5.Therapeutic Effect of High Frequency Oscillatory Ventilation on Premature Infants with Respiratory Distress Syndrome
yan, WANG ; xiang-yong, KONG ; zhi-chun, FENG
Journal of Applied Clinical Pediatrics 2006;0(14):-
Objective To evaluate the effect of the high frequency oscillatory ventilation(HFOV) on the treatment of premature neonates with respiratory distress syndrome(RDS).Methods Seventy premature infants with RDS were randomly grouped as HFOV group(n=33) and conventional mandatory ventilation group(CMV group,n=37),based on their fetal age,weight,age,and their clinical condition from Jan.to Sep.in 2009.The blood gas analysis was detected and compared between the 2 groups.Results In HFOV group,the inhaled oxygen concentration,pa(CO2) decreased after treatment for 6 h,which were significantly lower than those at the beginning of the therapy(Pa
6.Antibody to Ro-52 in patients with autoimmune liver disease
Dantong ZHAO ; Huiping YAN ; Yufen TAN ; Yan LIU ; Yan ZHAO ; Xia FENG ; Daijun XIANG
Chinese Journal of Microbiology and Immunology 2009;29(7):656-659
Objective To investigate the significance of antibody to Ro-52 in patients with autoim-mune liver disease(AILD). Methods One hundred and fifteen patients with abnormal liver functions, who had anti-Ro-52 detection by immunological blotting, were reviewed retrospectively. According to types of AILD, the clinical features were compared between patients with and without anti-Ro-52, respectively, κ test of concordance was used to provide a chance-corrected valve for immune-serological results. Results The rates of anti-Ro-52 in autoimmune hepatitis( AIH), primary sclerosing cholangitis(PBC) and AIH/PBC o-verlap syndrome groups were 32.43%, 24.56% and 33.33%, respectively, there were no significant differ-enees among three groups ( x2 = 0. 949, P >0. 05). The rate of anti-soluble hver antigen/liver-pancreas ( an-ti-SLA/LP) in AIH patients with anti-Ro-52 (58.33%) was higher than AIH patients without anti-Ro-52 ( 16.00% ,P < 0.05 ). The rate of anti-SLA/LP in AIH/PBC overlap syndrome patients with anti-Ro-52 (85.71%) was also higher than that of control group (28.57% ,P <0.05). Anti-Ro-52 and anti-SLA/LP had concordance according to κ test( κ >0.40, P <0.05). The average level of IgG in AIH/PBC overlap syndrome patients with anti-Ro-52 was higher than patients without anti-Ro-52 ( t = 2. 508, P < 0.05 ). Conclusion The rates of anti-Ro-52 in AIH, PBC and AIH/PBC overlap syndrome were of no significant differences. Anti-Ro-52 may have correlation with anti-SLA/LP. AIH/PBC overlap syndrome patients with anti-Ro-52 shewed higher IgG level than patients without anti-Ro-52.
7.Processing Technology Research of Fermentation and Purification of SUMO Protease UlP1
Xiu-Ping FENG ; Bai-Rong DU ; Dong-Mei YAN ; Xiang-Feng ZHAO ; Xun ZHU ;
China Biotechnology 2006;0(02):-
Nowadays,small peptides are always expressed in the form of fusion protein.The expression product contains many superfluous amino acids which can affect the biological functions of small peptides even expressed by GST fusion protein expression system.SUMO protease can cut SUMO fusion protein expressed by fusion expression system without any amino acid residues left on target protein thus become a hot topic in this field.Recombinant His-UlP1/pET3c/BL21(DE3)engineering strain was constructed by genetic engineering technology and the expression conditions were optimized in shake flaks.The process of high density fermentation was explored and different purification conditions were detected by chromatography.The results showed that SUMO protease could be expressed well after inducing the engineering strain by IPTG of 1.0mmol/L at 30℃ for 6 hours.The expression level of the strain in fermentation pots could reach 24.3% analyzed by SDS-PAGE.The purity of SUMO protease was more than 98% after further purification by cation exchange chromatography.The yield was 355mg SUMO protease per liter fermentation liquid.Western blot analysis demonstrated that there were immune reactions between IlP1 and 6?His antibodies,so it has established a good foundation for large-scale industralazation in the future.
8.VSMCs Proliferation and Mechanism of Signal Transduction by Angiotensin Ⅱ Type 1 Receptor Autoantibodies Mediated from Hypertensive Patients
Yan-Xiang SUN ; Yu-Hua LIAO ; Yu-Miao WEI ; Hai-Yan ZHANG ; Min WANG ; Feng ZHU ;
Chinese Journal of Hypertension 2006;0(09):-
Objective The autoantibodies against angiotensin Ⅱ type 1 receptor(AT_1 RAb)have been dis- covered in the patients with malignant hypertensive and preeclampsia,this autoantiboies(AT_1-AA)have an ago- nist-like activity effect similar to angiotensin Ⅱ(Ang Ⅱ).This study aimed at investigation the effect of Ang Ⅱ agonist-like activity by AT_1-AA on VSMCs proliferation was obtained from essential hypertensive patients. Methods VSMCs were cultured from aorta of WKY rats.The hypertensive patients" serum was purified by am- monium sulfate precipitation and affinity chromatography.The effect on VSMC proliferation this autoantilody was determined by BrdU incorporation.Total protein and the expression of phosphorylation JAK-STAT were assessed by Western blotting.Results AT_1RAb caused a significant increase in BrdU incorporation similar to Ang Ⅱ during 0-24 h reaching peak value at 12 h.The A value of in 450 nm was higher in AT_1RAb group (0.236?0.012)than AG490+AT_1RAb group(0.176?0.009),Losartan+AT_1RAb groups(0.119?0.006) and Serum Free group(0.127?0.006)(P
9.The cell-mediated immune status of patients with cytomegalovirus pneumonia after renal transplantation
Heli XIANG ; Wujun XUE ; Puxun TIAN ; Xiaoming DING ; Xiaoming PAN ; Hang YAN ; Jun HOU ; Xinshun FENG
Chinese Journal of Organ Transplantation 2011;32(10):592-595
Objective To evaluate the clinical value of adenosine triphosphate (ATP) determination in CD4+ cells in cytomegalovirus pneumonia after renal transplantation.Methods The ATP level of CD4+ T cells was measured by ImmuKnowTM kit.The ATP levels were determined in 187 renal transplant recipients before and 30,60,90,180 days after operation,and at the time of CMV pneumonia and 4 weeks after treatment of CMV pneumonia.The associations between ATP levels and CMV pneumonia were analyzed.Analysis of variance (ANOVA),Pearson-Spearman and relative risks were used for data analysis.Results 17 cases out of 187 renal transplant recipients were diagnosed as CMV pneumonia (9.1%),and the onset of CMV pneumonia started on the (2.8 ±1.2)month after renal transplantation.ATP concentrations in CD4+ T cells were significantly lower after operation than those before operation (P<0.01).ATP concentrations reached the lowest on the about postoperative day 90 (P<0.05),then increased gradually.In 17 recipients with CMV pneumonia,the ATP levels before and 30,90 days after operation,at the time of CMV pneumonia and 4th week after treatment of CMV pneumonia were (376 ±182),(283 ± 146),(196 ± 112),(145 ± 102) and (236 ± 117) μg/L respectively.ATP levels at the time of CMV pneumonia were significantly lower than any other time points (P<0.05).There was close correlation between ATP levels and CMV pneumonia.Conclusion The determination of ATP in CD4+ cells could reflect the status of cell-mediated immunity in renal transplant recipients,and could evaluate the severity and prognosis of CMV pneumonia and guide the clinical treatment.
10.Accuracy of serum S-100β protein and neuron specific enolase level in prediction of postoperative delirium in different age patients
Kaiyun FANG ; Yan ZHU ; Yaping FENG ; Xiang HE ; Jie SHANG ; Min ZHANG ; Wei ZHAO
Chinese Journal of Anesthesiology 2012;32(1):27-30
Objective To investigate the accuracy of serum S-100β protein and neuron specific enolase (NSE) level in predication of postoperative delirium (PD) in patients of different ages.Methods Four hundred ASA Ⅰ -Ⅳ patients of both sexes weighing 40-82 kg undergoing abdominal surgery performed under general anesthesia were divided into 4 age groups:group Ⅰ 18-44 yr; group Ⅱ 45-59 yr; group Ⅲ 60-74 yr and group Ⅳ ≥75 yr.The diagnosis of PD was made by using confusion assessment method.The incidence of PD was recorded within 72 h after operation.Each group was further divided into PD and non-PD subgroups.Blood samples were taken at 1 day before operation (T1),during their stay in PACU (T2) and at 24 and 72 h after operation (T3,4 ) for determination of serum S-100β protein and NSE concentrations.The receiver operating characteristic (ROC) curve for serum S-100β protein concentration in determining the PD efficacy was plotted.Results The incidence of PD was significantly higher and the duration was significantly longer in groups Ⅲ and Ⅳ than in groups Ⅰ and Ⅱ,and in group Ⅳ than in group Ⅲ (P < 0.05).There was no significant difference in the serum S-100β protein concentration between PD subgroup and non-PD subgroup in groups Ⅰ - Ⅲ ( P > 0.05).Compared with that at T1 and in nonPD subgroup,the serum S-100β protein concentration was significantly increased in PD subgroup in group Ⅳ,and the serum NSE concentration was significantly decreased at T2,3 in PD subgroup in group Ⅰ (P < 0.05).There was no significant difference in the serum NSE concentration between PD subgroup and non-PD subgroup in groups Ⅱ -Ⅳ.The analysis results of the ROC curve showed that:the area under the curve for the serum S-100β protein concentration and 95% confidence interval were 0.329 (0.127-0.531),0.352 (0.168-0.536),0.619 (0.466- 0.772) and 0.921 (0.846-0.995),the sensitivity was 50%,50%,56% and 88%,and the specificity was 29%,22%,46% and 86% in groups Ⅰ-Ⅳ respectively.Conclusion Increase in the serum S-100β protein concentration can be used in predicting the development of PD in patients ≥75 yr,but the serum NSE protein concentration can not be used.