Corneal thickness and morphology after orthokeratology of six-month lens wear among young Malay adults

@#Background: Menicon Z night orthokeratology (OK) lenses was introduced in Malaysia in 2015 and to date there is no report on its effects on the cornea. The objective of this study was to examine short term changes in corneal thickness and morphology of endothelial cells in young Malay adults after wearing Menicon Z night OK lenses. Methods: Corneal thickness was measured at the central and mid-peripheral locations of 20 participants aged 22.45±1.19 years using Tomey SP-3000 A-scan ultrasonography. Endothelial images of the central and peripheral locations captured using Tomey EM-3000 specular microscope were noted. Corneal thickness, endothelial cell density (ECD), coefficient of variation in cell size (CV), and hexagonality (HEX) at baseline, 24 hours, three months and six months after treatment were noted and analysed using repeated measure analysis of variance. Results: Central corneal thickness decreased significantly over a three-month period (p=0.001) and stabilised thereafter. There were no significant changes in thickness in all peripheral areas measured (p>0.05), and in ECD, CV and HEX after the six-month period (p>0.05). Conclusions: The current study showed that significant thinning of central cornea and none at the mid-periphery. OK lens wear with Menicon Z night lenses had no effects on corneal morphology over the six month period.

Hydroxysafflor yellow A attenuate lipopolysaccharide-induced endothelium inflammatory injury / 中国结合医学杂志

<p><b>OBJECTIVE</b>This study observed attenuating effect of hydroxysafflor yellow A (HSYA), an effective ingredient of aqueous extract of Carthamus tinctorius L, on lipopolysaccharide (LPS)-induced endothelium inflammatory injury.</p><p><b>METHODS</b>Eahy926 human endothelium cell (EC) line was used; thiazolyl blue tetrazolium bromide (MTT) test was assayed to observe the viability of EC; Luciferase reporter gene assay was applied to measure nuclear factor-κB (NF-κB) p65 subunit nuclear binding activity in EC; Western blot technology was used to monitor mitogen activated protein kinase (MAPKs) and NF-κB activation. Reverse transcription polymerase chain reaction (RT-PCR) method was applied to observe intercellular cell adhesion molecule-1 (ICAM-1) and E-selectin mRNA level; EC surface ICAM-1 expression was measured with flow cytometry and leukocyte adhesion to EC was assayed with Rose Bengal spectrophotometry technology.</p><p><b>RESULTS</b>HSYA protected EC viability against LPS-induced injury (P <0.05). LPS-induced NF-κB p65 subunit DNA binding (P <0.01) and nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor α (IκBα) phosphorylation was inhibited by HSYA. HSYA attenuated LPS triggered ICAM-1 and E-selectin mRNA levels elevation and phosphorylation of p38 MAPK or c-Jun N-terminal kinase MAPK. HSYA also inhibited LPS-induced cell surface ICAM-1 protein expression P <0.01) and leukocyte adhesion to EC (P <0.05).</p><p><b>CONCLUSION</b>HSYA is effective to protect LPS-induced high expression of endothelium adhesive molecule and inflammatory signal transduction.</p>

Study on Inhibitory Effects Mechanism of Scallop Skirt Glycosaminoglycan on Oxidative Stress Injury in Vein Endothelium Cells Induced by OX-LDL / 中国药房

OBJECTIVE:To investigate the inhibitory effects mechanism of scallop skirt glycosaminoglycan(SS-GAG)on inju-ry in human umbilical vein endothelium cells (HUVEC). METHODS:In the test,there was a negative control group,a model group and the groups of SS-GAG at high,middle and low concentrations(mass concentrations of 200,100 and 50 mg/L respective-ly). The cells in latter 3 groups were cultured in SS-GAG at different mass concentrations for 12 h,and then in 50 μmol/L oxidized low-density lipoprotein(OX-LDL)for 24 h. MTT method was used to detect cell viability and the activity of lactic dehydrogenase (LDH),the flow cytometer to determine the level of reactive oxygen species (ROS),real-time fluorescence quantitative poly-merase chain reaction (RT-PCR) to detect mRNA expression of lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1), and Western blot to detect NOX4 protein expression. RESULTS:Compared to the cells in the negative control group,those in the model group demonstrated lower viability,higher activity of LDH,higher level of ROS,and stronger expressions of LOX-1 mRNA and NOX4 protein. There was statistical significance (P<0.01). Compared to the cells in the model group,those in the groups of SS-GAG at high,middle and low concentrations showed higher viability,lower activity of LDH,lower level of ROS and weaker expressions of LOX-1 mRNA and NOX4 protein. There was statistical significance (P<0.01). CONCLUSIONS:SS-GAG can protect HUVEC to some degree by a mechanism which may be related to inhibiting ROS production via LOX-1/NOX4 pathway and relieving oxidative stress injury.

Morphologic observation of induced pluripotent stem cells induced by corneal endothelium cells with atomic force microscopy / 中华实验眼科杂志

Background Induced pluripotent stem cells (iPSCs)can differentiate into various types of somatic cells without causing ethical controversy and immune rejection in clinical activity,which is similar to differentiation ability of embryonic stem cells.So,iPSCs may be used as seed cells for tissue engineering corneal endothelial reconstruction.Objective The present study was to survey the morphologic change of iPSCs after coculture with corneal endothelium cells(CECs) under the atomic force microscopy(AFM).Methods Rabbit CECs and human MMC-iPSCs were isolated and cultured respectively.The iPSCs were identified with the marker by immunochemistry.iPSCs passaged for 7 days were then cultured with 60％ confluent CECs to establish the co-culture model.The surface morphology and cellular membrane ultrastructure of differentiated iPSCs after induced by CECs were examined by AFM combination with inverted microscope,and compared with CECs and undifferentiated iPSCs.Results Thelengthand width were(66.93±10.48)μm and (44.85 ± 8.14) μm in CECs,(12.51±1.40)μm and (10.93 ±1.69) μm in uninduced iPSCs,and(36.12±10.29) μm and(31.53±9.65)μm in CECs-induced iPSCs.Both the length and width values of CECs-induced iPSCs were statistically bigger than those uninduced iPSCs,with significant differences between them (P＜0.05),but no significant difference was seen in the width valne of CECs-induced iPSCs in comparison with CECs(P＞0.05).The convex structure of CECs cytomembrane surface showed the digitation in shape with the size and height(2.11 ± 1.03) μm and (115.68±92.08) nm respectively,and the concave structure of cytomembrane surface of CECs was fenestrae-like depression and the size was (1.49 ± 0.65) μm.The numerical valuc of mean square root roughness (Rq)and average roughness (Ra)of cytomembrane surface of CECs were(39.20±7.82)nm and (30.37±5.32)nm respectively.The convex surface of cytomembrane of iPSCs was granular-like in shape with size and height(0.39±0.22)μm and(13.11±9.18)nm respectively.The concave surface of cytomembrane of iPSCs was worm-eaten-like concave with the size(0.34±0.18)μm.The numerical value of Rq and Ra of geometrical parameters of cytomembrane surface of iPSCs were (26.60 ± 4.93)nm and (9.97 ± 3.78) nm respectively.The convex surface of cytomembrane of induced iPSCs was digital-like in shape with the size and height (1.91±0.76) μm and(106.55±77.27) nm respectively.The concave surface of cytomembrane of induced iPSCs was fenestrae-like depression and the size of concave was(1.6l±1.25) μm.The numerical value of Rq and Ra on surface of cytomembrane of induced iPSCs was (57.33± 12.80) nm and (43.63± 11.17) nm respectively.The numerical values of the size and height of convex,the size of concave,Rq and Ra on surface of cytomembrane in induced iPSCs were statistically bigger than in iPSCs(P＜0.05)and were not significant differences in comparison with CECs (P＞0.05).Conclusions Morphology of iPSCs translate toward the CECs after induce for 7 days under the AFM.This outcome lays the foundation for further study on iPSCs.

Central corneal thickness and corneal endothelium morphology in type 2 diabetes mellitus / 第三军医大学学报

Objective To study the changes of central corneal thickness(CCT) and corneal endothelium morphology in type 2 diabetes mellitus. Methods The CCT and corneal endothelium morphology in 60 diabetic patients(60 eyes) and 60 healthy volunteers(60 eyes) as the age and gender matched control group were examined by pachymetry and non contact specular microscopy. Stepwise regression analysis was performed to assess the effects of systemic factors on CCT and the corneal endothelial cell density of diabetic patients. Results Compared with that in the control group, the endothelial cell density decreased, and the coefficient of variation of cell area increased significantly( P 0 05). Compared to patients with non proliferative diabetic retinopathy(NPDR), the patients with proliferative diabetic retinopathy(PDR) had a significant reduction in cell density( P

Effect of curcumine on proliferation and heparanase expression in cultured blood vessel endothelium cells from ovarian cancer / 第三军医大学学报

Objective To explore the effect of curcumine on the proliferation and heparanase expression in blood vessel endothelium cells isolated from ovarian cancer tissues. Methods Blood vessel endothelium cells were obtained from pathologically identified ovarian cancer tissues by primary culture. The control group was the normal blood vessel endothelium cells. The experimental groups was treated with curcumine at 2,10,50 and 250 ?g/ml for 24,48 h and 72 h respectively. The cytostasis rate and cell cycle changes of the primarily cultured cells were detected by MTT assay and flow cytometry respectively. The transcriptional and translation levels of heparanase were detected by RT-PCR and immunofluorescent staining. The MDA content and LDH activity in the supernatant were detected too. Results In the curcumine groups,the cell proliferation was inhibited in a dose-dependent and time-dependent manner ( P

The effect of transcription factor SP1 decoy oligodeoxynucleotides on expression of ? Gal in SV-40-PED cells / 医学研究生学报

Objective:To investigate the role of transcription factor SP1 decoy oligodeoxynucleotides(ODN) on expression of ? Gal in SV-40-PED cells.Methods:Immortalized porcine aortic endothelial cells of the PED line were cultured and transfected with ?1,3galactosyltransferase(?1,3GT) specific decoy ODN.Cells transfected with mismatch ODN was used as negative controls.Twenty-six hours later the cells were collected.The expression of ? Gal was determined with fluorescence microscope and Western blot.The expression of ?1,3GT mRNA was examined by RT-PCR.Results:Fluorescence microscopy observed the decreased fluorescence of ? Gal after decoy ODN transfection.Western blot showed that the average absorbance of the PED cells transfected with decoy ODNs was(48.2?0.9).It is 52.6% of the mock group(P0.05).Conclusion:?1,3GT gene reduce actually occurs following transfection of decoy ODN.Porcine endothelial cells can be the targets of decoy ODN.

THE MORPHOLOGICAL CHARACTER ON LYMPHOCYTE HOMING AND THE EXPRESSION OF LFA-1,PECAM-1 IN THE RAT MESENTERIC LYMPH NODE / 解剖学报

Objective To study the morphological character of the high endothelial venules(HEVs) and the expression of lymphocyte function-associated antigen-1(LFA-1),platelet endothelium cell adhesion molecular-1(PECAM-1) in the rat mesenteric lymph node,inquire into their morphological representation when lymphocytes pass through the HEVs and the regulative action of LFA-l and PECAM-1. Methods To observe their morphological representation when lymphocyte pass through the HEVs wall and analyze the expression of LFA-1,PECAM-1 with the light microscope,electron microscope and immunohistochemistry method in the rat mesenteric lymph node. Results Lymphocytes contacted the endothelial cells by protuberances or membrane protuberances like welding spot,lymphocytes located in and between the endothelial cells or in the junction space of them in HEVs,there were lymphocytes in the lucid layer between inner and outer layers of basement membranes;LFA-1 mainly expressed at lymphocytes that adhered to endothelial cell in HEVs;PECAM-1 mainly expressed at the endothelial cells and the migrating lymphocytes.Conclusion 1.Lymphocytes contacted the endothelial cells by protuberances,then they got across the endothelial cells into the cellular junction space by the intraendothelial and interendothelial,and at last they emigrated from the basement membranes into the lymph tissue;2.LFA-1 involved in firm adheresion between lymphocytes and the endothelial cells;3.PECAM-1 probably had a relationship with that lymphocytes migrated endothelial cells.

Corneal Central Endothelial Cell Loss after Trabeculectomy with Mitomycin C

The effect of mitomycin on the corneal endothelial cell density was evaluated between the mitomycin group of 23 eyes who underwent trabeculectomy with mitomycin and the control group of 20 eyes who underwent trabeculectomy. The mitomycin(0.2 mg/ml) was applied under the scleral flap, Tenon's capsule or conjunctiva during trabeculectomy for 2-5 minutes(average: 4 minutes) and was irrigated with 50ml of balanced salt solution. Average central endothelial cell loss in the mitomycin group was 10.1 +/- 7.1% and 8.4 +/- 8.3% in the control group(P>0.05, t-test). A shallow anterior chamber with iridocorneal touch(Spaeth's grade 1 and 2) was developed in 12(52%) of 23 eyes in the mitomycin group and 9(45%) of 20 eyes in the control group(P>0.05, chi-square). The average central endothelial cell loss was 12.7 +/- 8.3% in 21 eyes with iridocorneal touch and 5.9 +/- 5.4% in 22 eyes that maintained their anterior chamber(P0.05, t-test). In 22 eyes that maintained their anterior chamber, average loss was 7.1 +/- 5.4% in the mitomycin group, and 4.8 +/- 5.4% in the control group(P>0.05, t-test). These findings suggest that intraoperative use of mitomycin(0.2 mg/ml) during trabeculectomy is unlikely to cause profound corneal endothelial cell loss and iridocorneal touch after the filtering surgery is associated with greater loss of endothelial cells.

Incubation of human ovarian cancer microvascular endothelial cells and formation of lumen-like structure in vitro / 第三军医大学学报

Objective To establish the magnetic activated cell sorting system(MACS)for isolation and incubation of human ovarian carcinoma-derived microvascular endothelial cells(ODMECs) and to observe their ability to form tubule-like structure(TLS) in vitro.Methods MACS was used to purify ODMECs with antiCD31 antibody and ODMECs were identified according to their morphological,biochemical and phenotypic characteristics.The ability of ODMECs to intake acetylated-low-density lipoprotein(DIL-acLDL) and Ulex europeaus agglutinin-1(UEA-Ⅰ) was assayed to observe the formation of TLS in vitro.Results Flow cytometry showed that the purity of ODMECs was up to 99.6%.The ODMECs were characterized by cobble stones,contact inhibition,expressed CD31,CD105 and FⅧ-Rag,and exhibited the ability to bind to UEA-1,intake DIL-acLDL and form TLS during incubation.Conclusion The MACS we established can be used to isolate and incubate human ODMECs and study the formation characteristics of TLS.

INFLUENCE OF ANGIOTENSIN CONVERTING ENZYME INHIBITORY PEPTIDES ON THE EXPRESSION OF eNOS,iNOS,ET-1 mRNA IN HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS / 营养学报

Objective and Method: To study the influence of angiotensin converting enzyme inhibitory peptides (ACEIP) on the expression of eNOS, iNOS, ET-1 mRNA in cultured human umbilical vein endothelial cells by semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) method and then investigate its mechanism. Results: Campared with the control, the expression of ET-1 and iNOS was lower and the expression of eNOS was higer in different ACEIP groups. Conclusion: The antihyertensive function of ACEIP partly depended on the its effect to lessen the expression of ET-1,iNOS and induce the expression of eNOS in cultured human umbilical vascular endothelial cells.