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.Effects of isoflurane or sevoflurane combined with remifentanil anesthesia on blood amyloid beta protein in elderly patients undergoing abdominal surgery
Kaiyun FANG ; Xiang HE ; Yan ZHU ; Yaping FENG ; Chunling LI ; Yiming REN ; Li TAN
Chinese Journal of Anesthesiology 2014;34(10):1178-1180
Objective To compare the effects of isoflurane or sevoflurane in combination with remifentanil anesthesia on blood amyloid beta protein (Aβ) in the elderly patients undergoing abdominal surgery.Methods Two hundred patients of both sexes,aged 65-75 yr,weighing 51-76 kg,of ASA physical status Ⅰ or Ⅱ,scheduled for elective abdominal surgery under general anesthesia,were randomly divided into 2 groups (n =100 each) using a random number table:isoflurane combined with remifentanil anesthesia group (IR group) and sevoflurane combined with remifentanil anesthesia group (SR group).Fifty healthy elderly subjects served as control group (group C).After anesthesia was induced with iv penehyclidine,sufentanil,propofol and vecuronium,the patients were endotracheally intubated and mechanically ventilated.In group IR,anesthesia was maintained with inhalation of isoflurane (end-tidal concentration 1.68 %,in IR group) or sevoflurane (end-tidal concentration 1.71%,in SR group),and target-controlled infusion of remifentanil (target plasma concentration 2-6 ng/ml).At l day before surgery and 3 days after surgery,the patients' cognitive function was assessed using Mini-Mental State Examination (MMSE),the development of postoperative cognitive dysfunction (POCD) was recorded,and blood samples were taken for determination of serum Aβ40 and Aβ42 concentrations.Results The incidence of POCD was 5% (in C group),56% (in IR group) or 22% (in SR group),and there was no significant difference among the three groups.There were no significant differences in the serum Aβ42 and Aβ40 concentrations after surgery among the three groups.Conclusion The mechanism by which sevoflurane or isoflurane in combination with remifentanil anesthesia results in POCD is not related to the levels of blood Aβ40 or Aβ42 in the elderly patients undergoing abdominal surgery.
10.The effects of inner vertical outer spiral complex scaffold in repairing long segment of urethral defect
Xiaowen DU ; Haohao CHEN ; Qing LIU ; Jianyang XIANG ; Qiao WANG ; Ting XU ; Qiuliang YAN ; Chao FENG
Chinese Journal of Urology 2017;38(1):59-65
Objective To explore the possibility of the inner vertical outer spiral complex tubular urethra scaffold vascularization in repairing long segment of urethral defect.Methods From August 2014 to October 2015,27 clean male New Zealand white rabbits were divided into 3 groups,S1 group was transfected recombinant vascular endothelial growth factor(VEGF) gene lentiviral vector group.S2 group was vascular pedicle transfer tube group.C group was simple stent group.A 3.0 cm inner vertical outer spiral complex scaffold was constructed by using the small intestine acellular matrix (SIS) and polylactic acid copolymer (PLGA) modified by type Ⅰ collagen surface,and adipose-derived mesenchymal stem cells (ADSC) and smooth muscle cells after transformation from New Zealand white rabbits.In S1 group,the seed cells were transfected by recombinant vascular endothelial growth factor (VEGF) gene lentivirus,which express VEGF protein.The complex scaffold was used to repair 3.0 cm rabbit urethral defect In S2 group,the untransfected cells were seeded into the scaffold and embedded in the skin near the groin artery 3 weeks for repairing urethral defect with vascular pedicle transfer tube.In group C,the unseeded scaffold was used to repair the urethral defect alone.Postoperative observation and urethrography were followed 4,8 and 24 weeks after implantation.The HE staining,fluorescence tracing,immunohistochemical and scanning electron microscopy were evaluated at the same phase.Results In S1 group,there were one urinary fistula and one urethral stricture-related death,respectively.The urethra was smooth and patent,histological examination showed active hyperplasia of urethral capillary.In S2 group,there were one urinary fistula and two urethral stricture-related deaths,respectively.The urethral was rough,local thinning or dilated.Fat accumulation and mucosal contraction were found in the urethral submucosal,respctively.In C group,there were one urinary fistula,three hypospadias,and three urethral stricture-related deaths.The thickness of the urethra was uneven and stricture bending.The urethral mucosa was poorly repaired and the scar was narrow.HE and CD31 staining showed that S1 and S2 groups were active in the proliferation of urethral capillaries,and the angiogenesis was abundant.VEGF staining showed that the cytoplasm of endothelial cell layer,smooth muscle layer of vascular wall and the urothelial epithelial cell layer were fully expressed at 24 weeks,especially in epithelial cell layer.CKpan staining showed that the epithelium of S1 and S2 group developed to stratified epithelium,and the morphology of urethra was similar to normal urethra at 24 weeks.The urethral epithelial in C group of grew poor as single-level,irregular arrangement,24 weeks is still a lack of effective stratified epithelium.HE and oα-SMA staining showed that the smooth muscle and actin gradually increased in group S1 and S2,α-SMA staining in group C was scarce and increased at 24 weeks.PLGA was encapsulated by the surrounding tissue and the structure of electrospinning was clear after 4 weeks,absorbed and degraded after 8 weeks and absorbed after 24 weeks.Conclusions The inner vertical outer spiral comnplex tubular urethra scaffold maybe a reasonable method in repairing long segment urethral defects,and the methods of tubular urethra scaffold vascularization by transfected VEGF gene recombinant lentiviral vector and vascular flap deserve more research.