1.Pharmacogenetic research in the association between human leukocyte antigen and adverse drug reactions
Yan XIONG ; Wei ZHANG ; Xiaoping CHEN
Journal of Central South University(Medical Sciences) 2014;(7):749-757
With the rapid development of pharmacogenetics, more and more studies have shown evidence in the association between polymorphisms at the human leukocyte antigen (HLA) loci and severe adverse drug reactions (SADRs). Several HLA-B alleles proved to be associated with SADRs for drugs such as carbamazepine, allopurinol, lamotrigine, and lfucloxacillin. hTe USA Food and Drug Administration (FDA) has even recommended routine screening for HLA-B allele before the use of abacavir and carbamazepine. With the completion of human genome project and the Hapmapproject, several new pharmacogenetics approaches such as genome-wide association study (GWAS) have emerged. hTese newly developed methods will undoubtedly accelerate the identiifcation and clinical utilization of the pharmacogenetic biomakers. In addition, the immunogenetic mechanisms by which the HLA alleles cause SADRs are explored at the cellular and molecular level. hTis review focuses on the recent progresses in HLA alleles and ADRs regarding both the clinical translation and modern pharmacogenetic methods.
2.Soluble expression and characterization of mouse lactate dehydrogenase-C4 in Escherichia coli
Duo ZHANG ; Yongzhong XIONG ; Ping CHEN
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
Objective A prokaryotic expression vector was constructed by inserting the coding sequence of mouse sperm specific lactate dehydrogenase C into pET-28a(+)and the recombinant mLDHC44 protein was purified by Ni+-NTA agrose.Methods The cDNA of mouse sperm specific lactate dehydrogenase C was obtained by RT-PCR,with total RNA of mouse testis tissues as templates.The coding sequence of mouse LDHC4 was amplified by PCR with specific primers.This recombinant vector was transformed into Escherichia coli BL21(DE3).The recombinant mLDHC4 protein was induced by isopropy-?-D-thiogalactoside(IPTG)and identified by sodium dodecyl sulfate polyacrylamide electrophoresis and LDH activity determination.After purified with Ni+-NTA agrose,the mLDHC4 protein was probed with antisera from the pVAX1-mLDHC4 vaccine(the eukaryotic expression vector of mouse sperm specific lactate dehydrogenase C)immunized BALB/c mice by Western blot analysis.Results After digested with BamH I-EcoR I,the recombinant plasmids produced right fragment which was about 1000bp.Sequencing showed that the sequence of the cloned fragment was in agreement with sequence in GenBank.This recombinant vector was named as pET-28a(+)-mLDHC4.With induction of IPTG,The recombinant protein with molecular weight of about 35 kD was expressed and the enzyme activity of this protein was high.After purified with Ni+-NTA agrose,this mLDHC4 protein formed a specific band by sodium dodecyl sulfate polyacrylamide electrophoresis and probed with antisera from immunized BALB/c mice and then formed a specific band in the nitrocellulose membrane.Conclusion The coding sequence of mouse lactate dehydrogenase subunit C had been cloned into the prokaryotic expression vector pET-28a(+)and the mLDHC4 protein could be expressed at a high level,the specificity of this protein was high and the activity was strong.
3.Comparison and analyse FAB classification of myelodysplastic syndrome with WHO classification of myelodysplastic syndrome
Jing ZHANG ; Shumin XIONG ; Bing CHEN
Chinese Journal of Postgraduates of Medicine 2006;0(03):-
Objective By compared FAB classification of myelodysplastic syndrome with WHO classification of myelodysplastic syndrome,understanded the proposed WHO new classification of myelodysplastic syndrome as soon as possible.Methods Retraced and analysed 78 cases of myelodysplastic syndrome based on examination of the bone marrow.Restults(1)According to the FAB classification,divided myelodysplastic syndrome into 5 kinds:RA,RARS,RAEB,CMML,RAEB-T.(2)According to the WHO classification,its subtypes were adjusted again,and its concluded:RA,RARS,RCMD,RAEB-Ⅰ,RAEB-Ⅱ,5q-del.(3)There were 9 cases possess chromosome to revise particularly in the chromosome check 28 cases myelodysplastic syndrome. Conclusion WHO classification has clinical guiding significance for early diagnosis,therapy observation and prognosis decision.
4.Vesicoureteral reflux caused by benign prostatic hyperplasia
Weimin CHEN ; Lishen XIONG ; Qing ZHANG
Chinese Journal of Urology 2001;0(06):-
Objective To study the correlation between benign prostatic hyperplasia (BPH) and vesicoureteral reflux (VUR). Methods Intravenous radionuclide cystography (IVRC) was performed on 88 BPH patients and micturition cystourethrography on 30 to find if there was any VUR and the degree of reflux. Results 31 patients had VUR of which there were 6 grade Ⅰ,7 grade Ⅱ,6 grade Ⅲ,6 grade Ⅳ and 6 grade Ⅴ.9 were on the right,6 on the left and 16 bilateral. Conclusions VUR do occur in some BPH patients and IVRC is the better method for its diagnosis and evaluation.
5.Color Doppler imaging feature of localized Castleman disease
Luzeng CHEN ; Liangliang ZHANG ; Bin WANG ; Hui ZHANG ; Yan XIONG
Chinese Journal of Ultrasonography 2012;21(8):701-703
Objective To summary the color Doppler imaging feature of localized Castleman disease.Methods From January 1997 to November 2011,32 localized Castleman diseases which were proved by pathology were analyzed.Results Round-like,hypoechoic,hypervascular lesions were showed in 23 hyalinevascular type lesions,2 of them with calcium,3 of them with structure liked lymph node hilum.Round-like,hypoechoic,hypervascular lesions or normal lymph node were showed in 5 plasma- type lesions and 4 mixed type lesions.Conclusions When round like,hypoechoic,hypervascular lesion is found by ultrasonography,Castleman should be considered.Calcium or lymph nod hilum-like structure is special finding in diagnosis of Castleman disease by ultrasonography.
6.Prostaglandin E_2 inhibits transforming growth factor-?_1 triggered lung fibroblast-myofibroblast transdifferentiation and collagen Ⅰ production in vitro
Xiang LONG ; Shengdao XIONG ; Weining XIONG ; Yongjian XU ; Zhenxiang ZHANG ; Yong CAO ; Jun CHEN ; Xiaohong XU
Chinese Journal of Pathophysiology 1989;0(05):-
AIM:To observe the effects of prostaglandin E2 on transforming growth factor-?1(TGF-?1)triggered human lung fetus fibroblast(HLF)transdifferentiation and connective tissue growth factor(CTGF),collagen type I(COLⅠ)expression.METHODS:HLFs were treated with TGF-?1,the cells underwent phenotypic change to myofibroblast.The marker of myofibroblast-?-smooth muscle actin(?-SMA)was detected by immunofluorescence.The ?-SMA content was measured by Western blotting.The changes in CTGF and COL Ⅰ at transcription levels were estimated by RT-PCR method.CTGF protein expression was evaluated by immunocytochemical.Cell culture medium hydroxyproline amount was measured by colormetric assay.RESULTS:PGE2 blocked TGF-?1 induced ?-SMA positive myofibroblast transformation(P
7.Enhancement and assessment of the fundus image.
Mengmeng CHEN ; Xingliang XIONG ; Guang LI ; Tingting ZHANG
Journal of Biomedical Engineering 2014;31(5):1144-1148
A new enhancement method is proposed based on the characteristics of fundus images in this paper. Firstly, top-hat transform is utilized to weaken the background. Secondly, contrast limited adaptive histogram equalization (CLAHE) is performed to improve the uneven illumination. Finally, two-dimensional matched filters are designed to further enhance the contrast between blood vessels and background. The algorithm was tested in DIARETDB0 databases and showed good applicability for both normal and pathological fundus images. A new no-reference image quality assessment method was used to evaluate the enhancement methods objectively. The results demonstrated that the proposed method could effectively weaken the background, increase contrast, enhance details in the fundus images and improve the image quality greatly.
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Contrast Media
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Fundus Oculi
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Humans
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Image Enhancement
8.Effect of GABA on expression of VEGF and MMP-9 in monocrotaline-induced pulmonary hypertension rats
Gang CHEN ; Jun SONG ; Hong XIONG ; Lujie ZHANG ; Jinquan LV
Chongqing Medicine 2014;(30):4043-4045
Objective To study the therapeutical effect of γ-aminobutyric acid on monocrotaline(MCT )induced pulmonary hy-pertension rats ,and to elucidate the expression of VEGF and MMP-9 .Methods Thirty SD rats were randomly divided into 3 groups :a normal control group(control group) ,a MCT-induced pulmonary hypertension group(model control group) ,and anγ-ami-nobutyric acid treatment group(treatment group) .The mean right ventricular pressure(mRVP)were detected ,the right ventricular hypertrophy index(RVHI)were measured ,WT% and WA% were evaluated ,and the expression of VEGF mRNA in the lung tissue and MMP-9 were detected wtih FQ-PCR and immunohistochemical staining method respectively .Results mRVP ,RVHl ,WT% , WA% and the expression of MMP-9 and VEGF mRNA of treatment group were lower than those in the model control group(P<0 .05) ,but higher than the control group(P<0 .05) .Conclusion GABA has a therapeutic effect on pulmonary hypertension rats through regulating the expression of VEGF mRNA and MMP-9 protein .
9.Transcription factor ETS-1 mediates the expression of miRNA-21 induced by arsenic in human hepatic L-02 cells
Caihua QI ; Aihua ZHANG ; Xiong CHEN ; Baofei SUN
Chinese Journal of Endemiology 2017;36(1):26-31
Objective To investigate the effect of sodium arsenite (NaAsO2) on the expression of miRNA-21 (miR-21) mediated by transcription factor ETS-1 in human normal hepatocytes (L-02).Methods Dose-effect study:The L-02 cells were treated with different doses of NaAsO2 [0.0 (control),2.5,5.0,10.0,20.0,40.0 μmol/L] for 24 h.Time-effect study:L-02 cells were exposed to 0 (control) and 20 μmol/L NaAsO2 for 12,24,36 and 48 h (n =6).ETS-1 and miR-21 were treated with ETS-1 shRNA and miR-21 inhibitor,respectively.The cells treated with ETS-1 shRNA (100 nmol/L) were divided into 4 groups:①ETS-1 shRNA NC treatment alone (control group);②ETS-1 shRNA NC combined with NaAsO2 (20 μ,mol/L) treatment group;③ETS-1 shRNA treatment alone group;④Treatment with ETS-1 shRNA and NaAsO2 (20 μmol/L) group.The MiR-21 inhibitor (100 nmol/L) treated cells were also divided into 4 groups:① miR-21 inhibitor NC treatment (control group);② miR-21 inhibitor NC combined with NaAsO2 (20 μmol/L);③miR-21 inhibitor group;④miR-21 inhibitor combined with NaAsO2 (20 μ mol/L) treatment group.The expression of ETS-1 mRNA and miR-21 were detected by quantitative real-time PCR (qRT-PCR);the protein expression of ETS-1 was detected by Western blotting.Results Dose-effect study:The expression of ETS-1 mRNA in the groups of 0.0 (control),2.5,5.0,10.0,20.0 and 40.0 μmol/L was 1.008 ± 0.028,1.552 ± 0.029,1.697 ± 0.050,1.842 ± 0.077,2.233 ± 0.096 and 2.235 ± 0.092;miR-21 expression was 1.025 ± 0.094,1.552 ± 0.072,1.683 ± 0.066,1.915 ± 0.171,2.337 ± 0.195 and 2.592 ± 0.177;the expression of ETS-1 protein was 1.060 ± 0.045,1.267 ± 0.160,1.386 ± 0.087,1.723 ± 0.196,2.208 ± 0.122 and 2.284 ± 0.224,respectively,and the differences were statistically significant (F =47.797,8.959,65.748,all P < 0.05),the NaAsO2 dose groups were significantly higher than those of the control group (all P < 0.05),and there was a dose-effect relationship.Time-effect study:The expression of ETS-1 mRNA in L-02 cells was 1.253 ± 0.175,1.623 ± 0.220,1.771 ± 0.324 and 1.913 ± 0.251,respectively at 12,24,36 and 48 h;the expression of miR-21 was 1.502 ± 0.111,1.716 ± 0.113,1.979 ± 0.186 and 2.452 ± 0.304;the expression of ETS-1 protein was 1.196 ± 0.105,1.502 ± 0.076,1.651 ± 0.074 and 1.839 ± 0.139,respectively,there were significant differences between the groups (F =14.936,39.180,39.441,all P < 0.05).The expression of various time points of exposure to NaAsO2 was significantly higher than those in the control group (1.044 ± 0.115,1.044 ± 0.124,1.108 ± 0.088,1.053 ± 0.061;1.092 ± 0.061,1.096 ± 0.169,1.024 ± 0.111,1.057 ± 0.146;1.020 ± 0.017,1.049 ± 0.121,1.024 ± 0.089,1.031 ± 0.124,all P< 0.05),and there was a time-effect relationship.ETS-1 shRNA and miR-21 inhibitor treatment:compared with ETS-1 shRNA NC combined with NaAsO2 (20 μmol/L),ETS-1 shRNA and NaAsO2 (20 μmol/L) could significantly inhibit the expression of ETS-1 (0.912 ± 0.238 vs 1.641 ± 0.225,P < 0.05),and down-regulated the expression of miR-21 (1.313 ± 0.334 vs 2.363 ± 0.252,P < 0.05).There was no significant difference of ETS-1 mRNA expression between miR-21 inhibitor and NaAsO2 (20.μmol/L) group (1.580 ± 0.077 vs 1.576 ± 0.065,P > 0.05) compared with miR-21 inhibitor NC and NaAsO2 (20 μmol/L).Conclusions The expression of ETS-1 and miR-21 in L-02 cells is significantly higher than those in control.The high expression of ETS-1 mediates NaAsO2-induced miR-21 overexpression,which may be an important molecular mechanism of arsenic-induced expression dysregulation of human hepatic miRNAs and liver damage.
10.Effects of hydrotalcite on healing quality of active gastric ulcer with Helicobacter pylori infection
Min ZHANG ; Xiong CHEN ; Canxia XU ; Dinghua XIAO
Chinese Journal of Digestion 2015;35(10):692-695
Objective To observe the effects of hydrotalcite on histological ulcer healing quality in patients with Helicobacter pylori (Hp) positive active gastric ulcer .Methods A total of 145 patients with H p positive active gastric ulcer were divided into two groups .The control group was treated with esomeprazole ,amoxicillin and furazolidone triple therapy .The treatment group was treated with above triple therapy and hydrotalcite .After four-week treatment ,gastroendoscopy was repeated .The sections of gastric biopsy specimens were stained with hematoxylin-eosin (HE) staining ,Van Gieson staining , Alcian blue-periodic acid stiff (AB-PAS ) staining , streptavidin-perosidase (SP ) immunohistochemistry staining and computer imaging analysis technology were applied to observe maturity type of mucosal structure at the margin of ulcer ,the content of collagen and neutral mucus ,and the changes of expression of epithelial growth factor receptor (EGFR) and basic fibrobast growth factor (bFGF) before and after treatment .Paired samples t test was performed for comparison before and after treatment .F test and Chi-square test were used for comparison between two groups .Results The percentage of maturity type of regenerated mucosal structure of treatment group was 62 .9% (39/62) ,however that of control group was 40 .6% (26/64) ,and the difference was statistically significant (χ2 =13 .09 , P=0 .03) .Compared with those before treatment , the content of collagen in granulation tissue and neutral mucus in regenerated mucosa increased in both groups after treatment , and the increase was more significant in treatment group ((55 .1 ± 10 .4)% and (45 .8 ± 7 .1)% ,respectively);and the differences were statistically significant (F= 12 .85 and 18 .17 ,both P<0 .01) .Compared with those before treatment ,the percentage of EGFR and bFGF positive cells of both two groups increased , and the increase was more significant in treatment group ((49 .5 ± 8 .4)% and (48 .8 ± 9 .4)% ,respectively) ,and the differences were statistically significant (F=12 .17 and 18 .73 ,both P<0 .01) .Conclusion Hydrotalcite combined with anti-H p triple therapy can improve the maturity degree of structure and function of regenerated mucosa at the margin of ulcer in patients with H p positive active gastric ulcer and then improve the healing quality of ulcer .