1.RPTC cells toxicity induced by cisplatin and anti-apoptosis mechanism of BCL-2
Leping FENG ; Wei QIAO ; Zheng DONG
Basic & Clinical Medicine 2006;0(02):-
Objective To explore the mechnism of toxicity induced by cisplatin on renal proximal tubular cells(RPTC) line and anti-apoptosis mechanism of BCL-2 with transfection of different mutant BCL-2 in vitro.MethodsRPTC was translocated with different mutants BCL-2(s).Cell apoptosis induced by cisplatin on RPTC was analyzed with confocal and flurencent microscope.The cell apoptosis was measured with Hoechst33258 after treatment with cisplatin.Results Different mutant BCL-2(BCL-acta,BCL-cb5)were translocated on mitochondrial and Endoplasmic Reticulum(ER) respectively.BCL-acta protected RPTC from apoptosis induced by cisplatin more easily than BCL-cb5 group in a time-dependant manner(P
2.The changes of multifocal electroretinogram of the affected and fellow eyes of the patients with central serous chorioretinopathy
Feng DONG ; Haihua ZHENG ; Jin LI
Chinese Journal of Ocular Fundus Diseases 1999;0(02):-
ObjectiveTo evaluate the retinal function with central serous chorioretinopathy (CSC) and the fellow eyes (mf-ERG).MethodsTwenty patients (40 eyes) with unilateral CSC, diagnosed by fluorescein angiography, were examined on each eye of the patients by mf-ERG multifocal electroretinography. The patients aged from 31 to 53 (13 men, 7 women; mean age, 41 years), and the CSC eyes had visual acuities of 0.2 to 0.8. None of the patients had significant abnormality in the fellow eyes with visual acuities of 0.8 to 1.5. Twenty-four healthy people (29 normal eyes with visual acuities of 1.0 and more and without any retinal diseases) were studied simultaneously as the normal control. The stimulus matrix of 61 hexagonal elements were grouped into 5 concentric rings from the fovea outward to 30? of the degree of visual field. The first-order responses of mf-ERG were recorded. The first negative wave (N 1-wave) and positive wave (P-wave) of the mf-ERG were measured. Amplitude intensities and latechies for the two configurations were calculated and compared.ResultsCompared with the control eyes, the responses were not only depressed severely in P 1- and N 1-wave amplitude intensities in the ring 1~3 in the CSC eyes with CSC (P0.05). There were no significant differences in N 1- and P 1-wave latencies in 5 concentric rings among the control eyes, the fellow eyes or the CSC eyes.ConclusionsCSC is a kind of macular disease involving both eyes. It may cause the decrease of retinal function in macular area of CSC eyes and also influence the retinal function in macular area of the fellow eyes. mf-ERG is a sensitive way in examining visual function.
3.Expression of Stromal Cell-Derived Factor-1,Chemokine Receptor 4 and Vascular Endotheliar Growth Factor in Children with Wilms Tumor
zheng-hua, DONG ; shu-feng, ZHANG
Journal of Applied Clinical Pediatrics 1992;0(05):-
ObjectiveTo investigate the role of vascular endotheliar growth factor(VEGF),chemokine receptor 4(CXCR4) and stromal cell-derived factor-1(SDF-1) in Wilms tumor as well as their relationship with clinical features by examining the expressions of VEGF,CXCR4 and SDF-1.MethodsThirty cases of Wilms tumor samples and 12 cases of adjacent kidney tissue were collected from the First and the Third Affiliated Hospital of Zhengzhou University from May 2003 to May 2008.Thirteen boys and 17 girls aging from 4 months to 7 years old of whom were 22 favorable histologic types and 8 unfavorable histologic types.By means of cytoplasm to dye buffy for masculine cell,staining intensity and percentage of positive tumor cells serve as the judgment criteria for being positive or negative.All tissues would be tested by hematoxylin and eosin stain at the same time.ResultsThe positive expression rate of VEGF,CXCR4 and SDF-1 in Wilms tumor were 63.33%,70.0% and 53.33%.Those in adjacent normal kidney tissue were 25.0%,8.3% and 16.7%.The 2 groups were statistically significant(Pa
4.Sequence analysis of p62~(dok) amino acid and cDNA clone
Changhua WANG ; Feng DONG ; Chuanren DONG ; Hanqiao ZHENG ; Hailu YANG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM:To analyse sequences of p62 dok amino acid and cDNA and to investigate p62 dok tyrosine phosphorylation and its relation with p21 ras GAP. METHODS:The purified p62 dok was extracted from CHO/IR cells. The peptide sequence of p62 dok was carried out on a high performance analyzer. PCR was performed with the primers designed from the sequence of p62 dok amino acid. Western blot and immunoprecipitation were used to identify tyrosine phosphorylation of p62 dok and the binding of p62 dok with p21 ras GAP. RESULTS:The p62 dok cDNA is a 1863 bp sequence and code 481 amino acid with 15 tyrosine residues and a putative pleckstrin homology domain. The p62 dok protein is rich in PxxP motif. The tyrosine-phosphorylated p62 dok can bind p21 ras GAP. CONCLUSION:Perhaps p62 dok is a new signaling molecule and play an important role in insulin signaling networks through RAS/MAPK pathway.
5.Sequence analysis of p62dok amino acid and cDNA clone
Changhua WANG ; Feng DONG ; Chuanren DONG ; Hanqiao ZHENG ; Hailu YANG
Chinese Journal of Pathophysiology 2001;17(5):407-410
AIM:To analyse sequences of p62dok amino acid and cDNA and to investigate p62dok tyrosine phosphorylation and its relation with p21ras GAP. METHODS:The purified p62dok was extracted from CHO/IR cells. The peptide sequence of p62dok was carried out on a high performance analyzer. PCR was performed with the primers designed from the sequence of p62dok amino acid. Western blot and immunoprecipitation were used to identify tyrosine phosphorylation of p62dok and the binding of p62dok with p21ras GAP. RESULTS:The p62dok cDNA is a 1863 bp sequence and code 481 amino acid with 15 tyrosine residues and a putative pleckstrin homology domain. The p62dok protein is rich in PxxP motif. The tyrosine-phosphorylated p62dok can bind p21ras GAP. CONCLUSION:Perhaps p62dok is a new signaling molecule and play an important role in insulin signaling networks through RAS/MAPK pathway.
6.The correlation between serum miR-21 and miR-34a expression and the efficacy of Flofox chemotherapy regimen in patients with advanced gastric carcinoma
Feng DONG ; Junjun MA ; Zhenghao CAI ; Xuan ZHAO ; Minhua ZHENG
Practical Oncology Journal 2017;31(2):147-151
Objective The objective of this study was to investigate the levels of serum miR-21 and miR-34a in patients with advanced gastric cancer before and after Folfox chemotherapy and its relationship with chemotherapy efficacy.Methods Forty-five patients with advanced gastric cancer who underwent Folfox chemotherapy were enrolled from May 2015 to October 2015.The levels of miR-21 and miR-34a were analyzed by Real-time PCR before and after chemotherapy.The relationship between miR-21 and miR-34a levels and chemotherapy efficacy was analyzed.The predictive values of miR-21 and miR-34a on the efficacy of chemotherapy were evaluated by the receiver operating curve(ROC).Results After chemotherapy,6 cases(13.33%)were relieved,10 cases(22.22%)were stable disease(SD),29 cases(64.44%)were disease progress(PD),the objective response rate(ORR)was 13.33%,the disease control rate(DCR)was 35.56%,the median survival time was 8.1 months.The level of miR-21 was down-regulated and up-regulation for miR-34a after chemotherapy.They were statistically significant(P<0.05).The level of miR-21 in the chemotherapy group was lower than that in the ineffective group,and the level of miR-34a was higher than that of the ineffective group(P<0.05).MiR-21 was negatively correlated with the efficacy of chemotherapy(rs=-0.431,P<0.05),while miR-34a was positively correlated with it(rs=0.504,P<0.05).The median survival time was 7.6 months for low-level patients with miR-21 and 6.0 months for high-level patients.The median survival time of miR-34a patients was 5.5 months,compared with 8.3 months for high-level patients,with a statistically significant difference(P<0.05).The Log-Rank test showed that the median survival of patients with low miR-21 was higher than that of patients with high miR-21(P<0.05).The median survival of patients with high miR-34a was higher than that of patients with low miR-34a(P<0.05).The miR-21 combined with miR-34a in predicting chemotherapy efficacy under the curve(AUC)was 0.865,the sensitivity was 80.9% and the specificity was 89.8%.Conclusion miR-21 and miR-34a are correlated with chemotherapy efficacy median survival time of patients with advanced gastric cancer.
8.Construction of eukaryotic expression plasmid of bFGF gene in rats and its expression in tenocytes.
Yong, FENG ; Dong, ZHENG ; Shuhua, YANG ; Jing, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(1):27-30
The bFGF plays an important role in embryonic development of tendons and ligaments and in the healing of injuried tendons and ligaments. The eukaryotic expression plasmid of rat basic fibroblast growth factor (bFGF) gene was constructed in order to further investigate the bFGF function in molecular regulatory mechanism in the repair of tendons and ligaments and to provide the foundation for the clinical application. The cDNA fragments of bFGF were cloned from the skin of rats by RT-PCR, and recombinated to the pMD18-T vector. The cDNA encoding bFGF was cloned from the pMD18-T vector by RT-PCR, digested with restriction enzyme EcoR I Pst I and bound to eukaryotic expression plasmid pIRES2-EGFP to construct eukaryotic expression plasmid pIRES2-EGFP-bFGF. The pIRES2-EGFP-bFGF was transfected into the tenocytes by lipid-mediated ransfection technique. MTT test was used to detect the biological activity of bFGF in supernatants after the transfection. The expression of type I and III collagen genes was detected by using RT-PCR. It was verified that the pIRES2-EGFP-bFGF was successfully constructed, and its transfection into tenocytes could significantly enhance the biological activity of bFGF, and increase the expression of type I and III collagen mRNA, suggesting that pIRES2-EGFP-mediated bFGF gene therapy was beneficial to the repair of tendons and ligaments.
9.Follow-up and retreatment strategies in the anti-vascular endothelial growth factor-A therapy for neovascularization of age-related macular degeneration
Xiao-dong, SUN ; Zheng-yu, SONG ; Feng-hua, WANG
Chinese Journal of Experimental Ophthalmology 2012;30(5):385-387
Wet age-related macular degeneration (AMD) is one of major causes of blindness in elder people.Intraocular injection of anti-vascular endothelial growth factor (VEGF) -A regimen has made big breakthrough for the treatment on choroidal neovascularization of wet AM D,while long-term follow-up and necessary retreatments are the key issues to remain obtained visual acuity.Multiple strategies of wet AMD have been used in following-up and retreating based on the visual acuity,optical coherence tomography (OCT),ophthalmoscope and fluorescein fundus angiography (FFA) in abroad.However,there also are major differences in the patient' s composition,treatment habits and distribution of medical sources in China from Western.So we suggest to standardize the follow-up and retreatment strategies about intravitreal injection of VEGF-A for wet AMD as to achieve a better effectiveness.OCT-guided individual follow-up and retreatment strategies should be very helpful for maintaining a long-term efficacy,minimizing the treatment time and reducing medical cost.