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.Early effect of induced membrane technique for the reconstruction of chronic osteomyelitis defects in limbs of adult patients.
Bing XIE ; Jing TIAN ; Yan-feng JING ; Da-peng ZHOU ; Liang-bi XIANG
China Journal of Orthopaedics and Traumatology 2015;28(1):43-47
OBJECTIVETo investigate the early clinical efficacy of induced membrane technique for reconstruction of large bone defects after debridement in adults with chronic osteomyelitis of limbs.
METHODSFrom March 2010 to March 2012,a total of 23 adult patients with chronic osteomyelitis of limbs were treated in our department. There were 15 males and 8 females, with a mean age 35.2 years old (ranged from 26 to 49 years old). Sixteen patients had open fracture history. According to the lesion site, there were 12 cases of tibia, 7 cases of femur, 3 cases of humerus, and 1 case of both radius and ulna. Among them, 19 patients had diseases in diaphysis and 4 patients in the metaphysis. The mean interval from infection to operation was 6.9 months (ranged from 4 to 13 months). All the patients were treated by using induced membrane technique. The follow-up evaluation included clinical complications, time of bone healing and limbs function. The Chinese version of SF-36 scores was used in the assessment of quality of life pre- and post-operation.
RESULTSThe average duration of follow-up was (27.6 ± 5.3) months (ranged from 18 to 43 months). Two patients had postoperative flap edge necrosis, 1 patient had superficial iliac incision infection, no obvious complications were recorded. Twenty patients obtained radiological union at a mean time of 4.6 months (ranged from 3 to 7 months). Among them, 16 patients treated with lower limbs surgery achieved full weight-bearing at about 5.2 months (ranged from 4 to 8 months) postoperatively. Four patients suffered from reinfection during follow-up, but 3 of them achieved complete bone healing after the second surgeries with induced membrane technique. At the final follow-up, there was a substantial improvement in each dimension scores and total scores of SF-36 as compared with those before surgery.
CONCLUSIONWhen treating with adult chronic osteomyelitis of limbs, the induced membrane technique can effectively reconstruct large bone defects after debridement, significantly shorten treatment cycle, provide satisfactory results with minimal complications, promote good recovery of limbs function and require relatively simple operation technique.
Adult ; Chronic Disease ; Extremities ; surgery ; Female ; Humans ; Male ; Middle Aged ; Osteomyelitis ; surgery ; Reconstructive Surgical Procedures ; methods