2.Hypoxia-induced changes of retinal progenitor cells migration by chemotaxis factor 4
Ping-hong, LAI ; Ming-ying, LAI ; Shi-bo, TANG
Chinese Journal of Experimental Ophthalmology 2011;29(10):868-871
Background In vitro study showed that chemotaxis consist of chemotaxis factor 4(CXCR4)and stromal cells derived factor-1(SDF-1)and may play a role in the orientation and migration of retinal progenitor cells (RPCs)toward lesion.Overexpression of CXCR4 in RPCs can enhance the chemotaxis activity.Objective This work was to explore the feasibility and underlying mechanism of up-regulation of CXCR4 on RPCs induced by hypoxia.Methods RPCs were retained in an incubator with normal O2volume(16%)or hypoxia condition(10% O2)for 12 hours and 24 hours respectively.Flow cytometer cell analysis screening(FACS)was conduced to measure the proportion of CXCR4-expressing cells,and CXCR4,HIF-1 mRNA were analyzed by reverse transcription-polymerse chain reaction(RT-PCR).The chemotical effect of 30 mg/L SDF-1 to RPCs cultured under the hypoxia condition was assessed using Boyden chamber.Results The expression level of CXCR4(CXCR4 mRNA/β-actin mRNA)inRPCs cultured by 10% O2 for 12 and 24 hours were 0.28+0.07and 0.48+0.17 and increased by 1.75 and 3.00 fold more than that of 16% O2 culture group(0.16+0.02)(P<0.01).The expression level of HIF-1 mRNA(HIF-1 mRNA/β-actin mRNA)in RPCs cultured by 10% O2 for 12 and 24 hours were 0.18 ±0.07and 0.38 ±0.13 and increased by 3.00 and 6.30 fold more than that of 16% O2 culture group(0.06±0.01)(P<0.01).The chemotical effect of 30 μg/L SDF-1 to RPCs increased from 13.00% in 16% O2 culture group to 36.00% and 46.00% in the cells cultured by 10% O2for 12 and 24 hours.FACS revealed that the proportion of CXCR4+ cells in hypoxia-exposure for 12 and 24 hours were 26.90% and 46.10%,respectively,but that in 16% O2 culture group was 9.10%,showing a statistically significant difference(P < 0.01).Conclusions RPCs induced by hypoxia can enhance the expression of CXCR4 in RPE cells and the chemotaxia to SDF-1.The overexpression of H1F-1 in RPCs may be involved in the up-regulation of CXCR4 expression.
4.Study on relationship between helicobacter pylori IgG antibody components and coronary heart disease.
Yuqiong LAI ; Xili YANG ; Ming LI
Chinese Journal of Practical Internal Medicine 2001;0(03):-
Objective To probe the relationship between Helicobacter py lori(Hp) IgG antibody components and coronary heart disease(CHD). Meth ods Hp serum specif ic IgG antibodies (Hp-IgG) and antibody components were measured by immunoblot ( IBT) methods in 209 CHD patients and 191 healthy controls from Mar.2001 to Sep 2 003. Results Of the 209 patients with CHD, 152(73%) were serop ositive to Hp comp ared with 113(59%) of the 191 healthy controls(P=0.0042). The Hp antibody co n tained different antibody components, such as cytotoxin-associated gene product A (CagA), vacuolating cytotoxin protein A (VacA), urease A (UreA), urease B (UreB ) etc,and only seropositivity to component UreB66 in CHD group was significantl y higher than that in healthy control (P=0.0001). Multiple Logistic Regression analysis revealed that the Hp antibody component UreB66 was significantly correl ated with CHD (P
5.The effect of chloride on cultured rat ventricular myocytes during anoxia reoxygenation injury
Jie CHEN ; Ming HE ; Zhongfang LAI
Chinese Pharmacological Bulletin 1986;0(04):-
Aim To investigate the effects of chloride on the injury of the ventricular myocytes from anoxia-reoxygenation. Methods Under the conditions of anoxia-reoxygenation injury, primary cultured rat ventricular myocytes were treated with 4-acetanide- 4′-isothiocya- natostilbene -2,2′-disulfonic acid (SITS),4,4′,-dii sothiocya-nostilbene-2,2′-disulfonicacid (DIDS) or replaced Cl~- with equimolar gluconate, respectively. Then the cell viability and the contents of Lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD)and glutathione peroxidase (GSH-Px) in the media were measured. Results The release of LDH and MDA was significantly increased in the anoxia-reoxygenation group, while the cell viability and the activity of SOD, GSH-Px decreased significantly compared with those in the control group. In both Cl~--free+ A-R group and SITS+A-R group, LDH and MDA release was noticeably lower than those of the A-R group, while the cell viability and the activity of SOD, GSH-Px significantly increased compared with those in the anoxia-reoxygenation group. But the cell viability and the contents of LDH, MDA, SOD and GSH-Px in the DIDS+A-R group had no significant change compared with those in the anoxia-reoxygenation group.Conclusion Cl~- plays an important role in anoxia reoxygenation injury. SITS provides effective protection to the cardiac myocyte subjected to anoxia reoxygenation injury, while DIDS cannot provide such protection.
7.Analysis of Lymhocyte Subsets and Humoral Immunity Condition in Children with Kawasaki Disease Complicated with Coronary Artery Dilatation
wei, WANG ; jian-ming, LAI ; zhe-wei, LIU
Journal of Applied Clinical Pediatrics 2006;0(21):-
Objective To explore the clinical diagnosis,medication,curative effect,and prognosis of lymhocyte subsets and humoral immunity in children with Kawasaki disease(KD)complicated with coronary artery dilatation.Methods One hundred and seventy-one children(age of 4 months to 9 years)from Jun.2000 to Dec.2007 with KD in Capital Institute of Pediatrics were taken as research objective,65 healthy children(age of 5 months to 7 years)were taken as controls,the difference of the level variation of lymhocyte subsets and humoral immunity was detected and analyzed between them.The flow cytometry and scattering turbidimetry method were used to detect the numbers of T cell subsets [CD3+,CD3+CD4+,CD3+CD8+,CD19+,natural killer(NK)cell] and levels of immune globulin(IgG,IgA,IgM)of the peripheral blood of 171 patients,compare them with those of the control group,and compare them between patients with and without coronary artery dilatation.Results Compared with control group,in the acute period of KD the levels of CD3+,CD3+CD4+ decreased significantly(t=0.01,0.02 Pa0.05);of all the 171 children patient,53 cases were coronary artery expansion patients.Compared with non-expansion patients,levels of CD4+/CD8+,CD3+CD8+,IgG of coronary artery expansion patients were significant different and had statistical difference(t=0.02,0.04,0.03 Pa
8.Correlation analysis of chronic renal failure patients with dry eye
Jian-Hua, ZHENG ; Shi-Jia, LAI ; Yong-Ming, SUN
International Eye Science 2015;(1):172-173
?AlM: To investigate the clinical characteristics and influencing factors of chronic renal failure ( CRF) patients with dry eye, and to provide clinical reference.?METHODS:Sixty-one cases (122 eyes) of patients with CRF ( CRF group ) and 61 cases ( 122 eyes ) of healthy persons ( control group) were carried out on Schirmer ▏test ( S▏t ) , break-up time of tear film ( BUT ) , corneal fluorescein staining ( FL) , test results of two groups were compared and related factors of dry eye in CRF patients were analyzed.?RESULTS: The results of S▏t and BUT in CRF group were lower than that in the control group (P<0. 05). The proportion of tear secretion reduce in CRF group ( S▏t<10mm/5min) was 49. 2% ( 60/122 ), which was higher than that in the control group ( 10. 0%, 12/122 ), the difference was statistically significant ( X2 = 45. 39, P <0. 05). The percentage of instability of tear film in CRF group (BUT≤10s) was 75. 4% (92/122), which was significantly higher than that in the control group (27. 0%, 33/122) (X2=57. 1, P<0. 05). The positive rate of corneal FL was 37. 7% (46/122), which was higher than that of the control group (10. 7%, 13/122), there was a statistically significant difference (X2= 24. 34, P<0. 05).?CONCLUSlON:CRF patients with a decrease in tear film stability and tear secretion are susceptible population to dry eye, clinically should be paid attention to the treatment.
9.Anti-angiogenic mechanism of cordycepin on rhesus macaque choroid - retinal endothelial cell line cultured in high glucose condition
Xiao-Li, ZHU ; Xiao-Feng, SUN ; Ming-Ying, LAI
International Eye Science 2016;16(7):1237-1241
AIM: To investigate the angiogenesis effect and protective mechanism of cordycepin on rhesus macaque choroid- retinal endothelial ( RF/ 6A) cell line cultured in high glucose condition.
METHODS: Cultured RF/ 6A cells were divided into normal control group, high glucose group and high glucose (HG) + different concentration cordycepin groups (HG+ 10μ g/ mL group, HG+ 50μ g/ mL group, HG+ 100μ g/mL group). The cell proliferation was assessed using cholecystokinin octapeptide dye after treated for 48h. The cell migration was investigated by a Transwell assay. The tube formation was measured on Matrigel. Furthermore, the impact of cordycepin on high glucose - induced activation of VEGF and VEGF receptor 2 (VEGFR-2) was tested by Western blot analysis.
RESULTS: Compared with normal control group, cell viability markedly increased in high glucose group ( P <0. 05). Cordycepin inhibited RF/ 6A cell proliferation in a dose- dependent fashion: 10. 2 ± 0. 9%, 23. 4 ± 1. 5% and 31. 1±1. 2% inhibition as the concentrations of cordycepin were 10, 50 and 100μ g/ mL, respectively. The difference had statistically significant (P<0. 05) compared with high glucose group. The number of cell migration were 55. 6±2. 70, 87. 4 ± 2. 40, 65. 4 ± 2. 7, 57. 8 ± 2. 38, 62. 4 ± 2. 77 in normal control group, high glucose group and HG+10μ g/mL group, HG + 50μ g/ mL group, HG + 100μ g/ mL group respectively. Migration of RF/ 6A conspicuously increased in high glucose group ( P < 0. 05) compared with normal control group; while showing a gradually reducing trend with the increase of cordycepin dose and a statistically significant difference compared with high glucose group (P<0. 05). The number of tube formation were 18. 7±2. 08, 25. 7 ± 1. 52, 19. 9 ± 1. 57, 16. 3 ± 2. 51, 5. 67 ± 1. 72 in the abovementioned group. Similarly showing a gradually reducing trend with the increase of cordycepin dose and a statistically significant difference with high glucose group (P< 0. 05). In addition, the number of tube formation of RF/ 6A in high glucose group significant increased compared with normal control group ( P < 0. 05 ). The expression of VEGF and VEGFR-2 dramaticlly increased in high glucose group vs normal control group, oppositely gradually decreased with the increase of cordycepin concentrations, and had a statistically significant difference vs high glucose group (P<0. 05).
CONCLUSION: Cordycepin can suppress the proliferation, migration and tubu formation of RF/ 6A in high glucose condition, might via inhibiting expression of VEGF and VEGFR-2.
10.Study on uncertainty of six enzymological reference methods
Ming CHEN ; Wenquan LAI ; Yingguo WANG ; Jingang CAI
International Journal of Laboratory Medicine 2015;(1):77-79
Objective To develop an uncertainty study model of the enzymological reference method to systematically research the uncertainty influencing factors of 6 enzymological reference methods ALT,AST,LDH,AMY,GGT and CK for determining the key factors influencing the reference method and better guiding the establishment and operation of the enzymological reference methods.Methods The uncertainty of the six enzymological reference methods was evaluated by the theoretical evaluation com-bined with the experiment design according to series of standards including JJF1059-1999 Evaluation and Expression of Uncertainty in Measurement,JJF1135-2005 Evaluation of Uncertainty in Chemical Analytical Measurement,CANA-GL06 Guidance on Evalua-ting the Uncertainty in Chemical Analysis and CNAS-CL33 Guidance on the Application of Testing and Calibration Laboratories Competence Accreditation Criteria in the Field of Clinical Enzymology Reference Measurement.Results The established enzymo-logical uncertainty study model was C=ΔAΔt · 1ε · 1L ·V 1 +V 2 +V SV ·fT ·f pH ·fλ·fkit ·fresult .Conclusion The uncertainty e-S valuation model is set up and successfully applied in the uncertainty evaluation of the six enzymological reference methods.