2.Interaction of E3 ligase HUWE1 and eukaryotic translation initiation factor eIF4E.
Jun-Ping ZHANG ; Ai-Juan XIA ; Rui-An XU
Acta Pharmaceutica Sinica 2014;49(11):1543-1546
To explore the regulation of eIF4E, we screened the protein interacting with eIF4E from human cDNA library by using yeast two-hybrid system. Several clones interacting with eIF4E were identified. One of them was homologous with HUWE1 (HECT, UBA and WWE domain containing 1, also named as ARF-BP1, HECTH9 or HUWE1). Cell co-immunoprecipitation showed that eIF4E could bind to HUWE1 in mammalian cells. We also found that HUWE1 bearing the HECT domain is necessary for its association with eIF4E.
Animals
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Eukaryotic Initiation Factor-4E
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metabolism
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Humans
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Ubiquitin-Protein Ligases
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metabolism
3.Rosiglitazone Inhibited Aldosterone-Induced Mesangial Cell Proliferation via Blocking Phosphatidyl Inositol 3-Kinase Activation
kang-kang, XU ; ai-hua, ZHANG ; gui-xia, DING
Journal of Applied Clinical Pediatrics 2006;0(17):-
Objective To explore the inhibitory effect of rosiglitazone of peroxisome proliferator-activated receptor-?(PPAR?) agonist on aldosterone-induced mesangial cell(MC) proliferation.Methods Mouse primary MC were cultured and treated with aldosterone(100 nmol/L) in the presence or absence of rosiglitazone(1.0,2.5,5.0,10.0 ?mol/L).The incorporation of 3H-thymidine(3H-TdR) and cell count were used as the measure of MC proliferation.Cyclin D1 and cyclin A expression,PI3K and Akt phosphorylation were determined by Western blot analysis.Results 1.Aldosterone induced MC proliferation,as assessed by 3H-TdR incorporation and cell number,which were increased by 2.46-and 2.14-fold,respectively,in aldosterone-treated cells.Aldosterone-induced MC proliferation was inhibited by PPAR? agonist rosiglitazone in dose-dependent manner in mouse MC.2.Aldosterone induced cyclin D1 and cyclin A expression.Rosiglitazone reduced aldosterone-induced cyclin D1 and cyclin A expression in dose-dependent manner.3.Aldosterone induced PI3K/Akt activation in dose-dependent manner,incubation with 100 nmol/L aldosterone for 60 min,phosphorylation PI3K and Akt expression increased by above 3.0-fold.4.PI3K inhibitor LY294002 and Akt inhibitor significantly inhibited aldosterone-induced cyclin D1 and cyclin A expression.5.Rosiglitazone significantly inhibited aldosterone-induced PI3K/Akt activation,10 ?mol/L rosiglitazone almost completely blocked aldosterone-induced PI3K/Akt activation.Conclusion Rosiglitazone can block aldosterone-induced MC proliferation via inhibition of PI3K/Akt activation.
4.Effect of SP600125 on AngⅡ-Induced Transforming Growth Factor-1 and Fibronectin Expression in Human Mesangial Cells
kang-kang, XU ; ai-hua, ZHANG ; gui-xia, DING
Journal of Applied Clinical Pediatrics 2006;0(23):-
Objective To investigate the effect of c-Jun N-terminal kinase(JNK) specific inhibitor SP600125 on Angiotensin Ⅱ(AngⅡ)-induced transforming growth factor-?1(TGF-?1) and fibronectin (FN) expression in human mesangial cells (MC).Methods Human MC were isolated and cultured in vitro and were treated with AngⅡ in the presence or absence of JNK specific inhibitor SP600125.The protein was isolated or the supernate of medium was collected at the end of experiment.JNK,extracellular signal-regulated kinase(ERK1/2),and p38 mitogen-activated protein kinase(MAPK) activity were determined by Western blot method.TGF-?1 and FN were determined by enzyme linked immunosorbent assay(ELISA).Results SP600125 inhibited AngⅡ-induced Ser63 phosphorylation of c-Jun in a concentration-dependent manner,and JNK activity was reduced by 75% at 10 ?mol/L and by 90% at 20 ?mol/L.SP600125 had no effect on AngⅡ-induced ERK1/2 and p38 activity.TGF-?1 and FN protein were constitutively produced in MC,and production was significantly stimulated for 8 to 48 h after addition of AngⅡ.Preincubation of cells with SP600125(20 ?mol/L) significantly inhibited AngⅡ-induced TGF-?1 and FN production during this time period.SP600125 inhibited AngⅡ-induced production of TGF-?1 and FN in a concentration-dependent manner.Conclusion SP600125 inhibited AngⅡ-induced JNK activation and TGF-?1 and FN expression in human MC and may serve as the novel approach for the treatment of patients with chronic kidney disease.
5.Effect of Fas/FasL pathway on fluoride-induced apoptosis in human neuroblastoma SH-SY5Y cells
Ba-yi, XU ; Zhi-xia, XU ; Tao, XIA ; Ping, HE ; Ping, GAO ; Wei-hong, HE ; Ai-guo, WANG
Chinese Journal of Endemiology 2008;27(5):479-483
Objective To explore the effect of Fas/FasL pathway on fluoride.induced apoptosis in hurnan neumbla8toma SH-SY5Y cells.Methods The cell survival rate,percentage of apoptosis,and mRNA expression levels of Fas and FasL were measured respectively after the SH-SY5Y cells were exposed to O(control),20,40,80 mg/L sodium nuoride(NaF)for 24 hours/n vitro.Furthermore,the changes of the percentage of apoptosis and mRNA expression levels of Fas and FasL in 40 mg/L NaF-treated groups incubated with activaling or neutralizing anti-Fas antibody(CH11 or ZB4)also observed respectively.Results Compared with the control group(100.00%), the cell surval rates in 40,80 mg/L NaF-treated groups[(84.63±2.57)%,(69.04±5.63)%]were significandy lower(P<0.01).The percentage of apoptosis in 40,80 mg/L NaF.treated groups[(8.54±1.95)%.(17.94±2.71)%]were higher(P<0.05)than thal in the control group[(3.32±1.33)%],and increased with the dose of NaF.NaF could up-regulate Fas and FasL mRNA expression,and increased the Fas/β-actin [40 ms/L group (0.94±0.51),80 mg/L group(0.99±0.12)]and FasL/β-actin[40 mg/L group(0.96±0.42),80 mg/L group(0.99±0.24)] ratio,compared with the control[Eas/β-actin(0.50±0.33),FasL/β-actin(0.58±0.23)],both the difference had 8tatistical significances (P<0.05).NaF and CH I 1 had a synergisfic effect on apoptosis and mRNA expression levels of Fas and FasLL(F=32.89,18.46,.14.69,P<0.01)while NaF and ZB4 had an antagonistic effect (F=5.73,24.26,10.17,P<0.05 or<0.01).Conclusion NaF exposure can cause apoptosis in SH-Y5Y cells,and the Fas/FasL pmhway may play an important role in NaF-induced apoptosis.
6.Correlation of TLR2 and TLR4 gene polymorphisms with the susceptibility and recurrence of condyloma acuminatum.
Ji-feng LIU ; Bin QU ; Xiang-dong WANG ; Qi WANG ; Xiao-xia ZHAO ; Ai-e XU
National Journal of Andrology 2015;21(8):708-712
OBJECTIVETo explore the correlation of the gene polymorphisms of Toll-like receptor 2 ( TLR2) and TLR4 with the susceptibility and recurrence of condyloma acuminatum (CA).
METHODSUsing Snapshot, we detected the gene polymorphisms of TLR2 597(T/C), 1350(T/C), 15607(A/G), and 2258(G/A) and TLR4 896(A/G) and 1196(C/T) in the peripheral blood of 140 CA patients and 105 HPV-negative controls. We made comparisons between the CA patients and controls as well as between the cases of recurrent CA and those of non-recurrence at 6 months after treatment.
RESULTSThere were 72, 48, and 20 cases of genotype TT, TC, and CC of TLR2 597 (T/C), respectively, in the CA patients, as compared with 71, 31, and 3 cases in the controls. The gene frequency of mutant C was 31. 43% in the patients, significantly higher than 17.62% in the controls (χ2 = 12.04, P < 0.01), and it was 38.68% in the recurrent cases, remarkably higher than 27.01% in the non-recurrent cases (χ2 = 4.16, P < 0.05). There were 74, 49, and 17 cases of genotype TT, TC, and CC of TLR2 1350( T/C), respectively, in the CA patients, as compared with 73, 29, and 3 cases in the controls. The gene frequency of mutant C was 29. 64% in the patients, significantly higher than 16. 67% in the controls (χ2 =11.05, P < 0.01), and it was 36.79% in the recurrent cases, markedly higher than 25. 29% in the non-recurrent cases (χ2 = 4.18, P < 0.05). There were 44, 66, and 30 cases of genotype AA, AG, and GG of TLR2 15607(A/G), respectively, in the CA patients, as compared with 26, 58, and 21 cases in the controls. There was no significant difference in the gene frequencies of mutant G between the two groups (χ2 = 0.33, P > 0.05). No mutant genes of TLR2 2508 (G/A) or TLR4 896(A/G) and 1196(C/ T) were detected in either the CA patients or the controls. Linkage disequilibrium analysis showed a tight linkage between TLR2 597 (T/C) and 1350(T/C) (D' = 1, r2 = 0.93).
CONCLUSIONTLR2 597(T/C) is tightly linked to 1350(T/C), which is correlated with both the susceptibility and the recurrence of condyloma acuminatum.
Aged ; Case-Control Studies ; Condylomata Acuminata ; genetics ; Gene Frequency ; Genetic Linkage ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Genetic ; Recurrence ; Toll-Like Receptor 2 ; genetics ; Toll-Like Receptor 4 ; genetics
7.The Mutagenic Effect on PHB Accumulation of Acidiphilium cryptum DX1-1
Ai-Ling XU ; Shuai ZHANG ; Yan-Fei ZHANG ; Li LI ; Yu YANG ; Jin-Lan XIA ;
Microbiology 2008;0(10):-
The strain Acidiphilium cryptum DX1-1 producing PHB was irradiated respectively by UV and Co60 to raise PHB production. The results indicated that the effect of UV better than using Co60. One strain of the UV mutagenized called UV60-3 has the highest PHB production yield, showing final PHB concentra- tion of 28.56 g/L, 1.45 times higher than that of original strain. FT-IR spectroscopy analysis shows that the polymers obtained from the strain DX1-1 have the same IR spectra of standard PHB. Further research about the best appropriate C/N ratio of the mutant was done. The optimum ratio of C/N was about 3.76, the final PHB concentration reaches to 30.57 g/L.
8.Surveillance of Antimicrobial Resistance in Staphylococcus spp and Enterococcus spp
Xiaoping XU ; Xia GAO ; Xiaoxia CHI ; Junhui GAI ; Hui AI ; Zhuocheng LI
Chinese Journal of Nosocomiology 2006;0(03):-
OBJECTIVE To analyze the resistant rates of Staphylococcus spp and Enterococcus spp isolated from clinical infections to antibiotics,and to provide reference method for effective control infections of Staphylococcus.METHODS The Staphylococcus spp and Enterococcus spp were identified with VITEK-32 automicrobiology system(AMS) and GPI card,drug resistance was detected with VITEK-32 AMS and GPS-107 card.Laboratory data were analyzed by WHONET-5 statistic software.RESULTS Among 1 445 Staphylococcus spp and Enterococcus spp strains isolated from clinical samples,330 strains(22.8%) were Staphylococcus aureus,872 strains(60.3%) were coagulase negative Staphylococcus,213 strains(14.7%) were Enterococcus faecalis,and 30 strains(2.1%) were E.faecium.From S.aureus 223 strains(67.6%) were MRSA,718 strains(82.3%) of coagulase negative Staphylococcus were MRCNS.The detectable rates of MRSA and MRCNS in 2004 were 75.3% and(82.3%,) which were higher than those in 2003(48.4% and 78.4%).Neither strains of S.aureus nor strains of coagulase negative Staphylococcus were found resistant to vancomycin.MRSA and MRCNS resistant rates were found(higher) than MSSA and MSCNS.From the isolated strains of E.faecalis in 2004,the resistance rates to(ciprofloxacin,) nitrofurantoin,gentamicin-500,levofloxacin,and penicillin G were found higher than that in 2003.(E.faecium)(resistant) rates were found significantly higher than E.faecalis.CONCLUSIONS Staphylococcus spp and Enterococcus spp are the main pathogens leading to clinical infections.The findings of these(surveillance)(studies) will enhance our knowledge regarding the problem of antimicrobial resistance and will serve as a basis for future policies and practice styles.
9.Relationship between myeloperoxidase and catalase genetic polymorphism and their activities with arsenic poisoning caused by coal-burning
Bing, LIANG ; Ai-hua, ZHANG ; Xu-guang, XI ; Bi-xia, ZHANG ; Xiao-xin, HUANG
Chinese Journal of Endemiology 2009;28(3):272-275
Objective To detect genetic polymorphism of myeloperoxidase (MPO) gene and catalase (CAT) gene and their activities, and to analyze their relationship with arsenic poisoning caused by coal-burning. Methods One hundred and thirty arsenic poisoning patients were chosen as case group in Jiaole Village, Xingren County, Guizhou Province(an endemic area). One hundred and forty healthy residents living in 13 km away were chosen as control group. Their blood was collected. Polymerase chain reaction-restriction fragment length polymorphism technique(PCR-RFLP) was used to detect polymorphism of MPO-463G/A and CAT-262C/T. Ultraviolet spectmphotometer method was used to detect myeloperoxidase activity. Chromatometry method was used to detect catalase activity. Results The genotype frequency of MPO-463G/A at GG, GA, AA site was 47.24%(60/127), 44.09%(56/127),8.67% (11/127) in case group and 42.34% (58/137),48.17% (66/137)1,9.49% (13/137) in control group, respectively. The difference between the two groups was not significant(χ2 = 0.642, P > 0.05). The genotype frequency of CAT-262C/T, at CC, CT, TT site was 65.60%(82/125),28.80%(36/125),5.60%(7/125) in case group and 76.51%(101/132), 18.94% (25/132) ,4.55% (6/132) in control group, respectively, without significant difference (χ2 =3.845, P>0.05). The relationship between polymorphism of MPO-463G/A and CAT-262C/T and the risk of arsenic poisoning was not found in this study(ORadj= 1.36, 95%CI: 0.74-2.50 for MPO; ORadj=1.35, 95%CI: 0.69-2.63 for CAT). The activities of MPO and CAT were (25.30±8.70)U/L and (2.80± 1.09)×103 U/L in case group, while (22.76±7.59)U/L and (3.90±1.01)×103U/L in control group with a significant difference(F=0.760 for MPO, F=0.855 for CAT, all P < 0.05). The genotype of MPO-463G/A and CAT-262C/T was not found to have relationship with the activities of MPO, CAT(F=1.312,2.822 for MPO; F= 0.151,0.036 for CAT, P>0.05). Conclusions Genetic polymorphism of MPO-463G/A and CAT-262C/T is not found to have relationship with arsenic poisoning. Arsenic can lead to the change of MPO and CAT activity, which, however, may not be affected by MPO-463G/A and CAT-262C/T polymorphism.
10.Relationship between intracellular calcium and reactive oxygen species in sodium fluoride-induced injury in human neuroblastoma SH-SY5Y cells
Zhi-xia, XU ; Ba-yi, XU ; Tao, XIA ; Ping, HE ; Ping, GAO ; Li-juan, GUO ; Qiang, NIU ; Nan, HUNAG ; Ai-guo, WANG
Chinese Journal of Endemiology 2009;28(2):126-129
Objective To explore the relationship between intracellular calcium levels ([Ca2+]1) and reactive oxygen species (ROS) in sodium fluoride (NaF)-induced injury in human neuroblastoma SH-SY5Y cells. Methods The levels of [Ca2+]1 and ROS were measured in different exposed times(0,3,6,12,18,24 h) respectively after SH-SY5Y cells were exposed to 40 mg/L NaF in vitro, and the optimal expose time was selected. Furthermore, the changes of [Ca2+]1, ROS and LDH levels in 40 mg/L NaF-treated groups incubated with 38.23 mg/L BAPTA-AM or 380.40 mg/L ethylene glycol-bis-(beta-aminoethyl ether)-N, N, N', N'-tetraacetic acid (EGTA) or 16.32 mg/L N-acetyl-L-cysteine(NAC) also observed at the optimal expose time(12 h), respectively. Results At 3,6,12,18 and 24 h, [Ca2+]1 level(5620.0±226.3,4775.5±85.6,3312.3±87.5, 3047.0±75.0,2717.0±66.5) was significantly increased, and so was the ROS level(4449.53±324.61,7463.07±117.43,20 227.33±178.04,8817.56±200.13, 7975.61±92.90) except at 3 h, compared with 0 h(2115.0±24.0,4098.01±21.22, all P<0.05). The levels of [Ca2+]1 and ROS reached the peak at 3 h and 12 h, respectively. [Ca2+]1 and LDH levels in NaF-treated group [3279.5±94.0, (1057.50±64.35)U/L], NaF+NAC treated group[ 3583.0±350.7, (561.02±85.50)U/L], NaF+EGTA treated groups[3701.5±157.7, (1074.50±86.97)U/L], and BAPTA-AM treated group[2766.5±38.9, (521.43±40.80)U/L] had increased, compared with the control[2022.5±118.1, (186.97±8.73)U/L], the difference being statistically significant (P<0.05). ROS levels in NaF-treated group (19 003.04±332.34), and NaF+EGTA treated group(19 170.12±95.46) was higher than that in the controls(4060.98±145.66), the difference being statistically significant (P<0.05). NaF and NAC had antagonistic effect on ROS and LDH levels (F=976.11,43.54,P<0.05). And NaF and BAPTA-AM had antagonistic effect on [Ca2+]1, ROS and LDH levels (F=15.65,1515.53,115.00, P<0.05). Conclusions NaF-related calcium is released from the site of intracellular calcium storage, which induces ROS production, both of them caused cytotoxicity and the increase of LDH level in human neuroblastoma SH-SY5Y cells.