Overwork Affects Extracellular Matrix of Arterial Vessel Wall in Rats / 中国医学科学院学报

Objective To explore the effect of overwork (OW) on extracellular matrix of arterial vessel wall in rats. Methods Random number grouping method was employed to assign 18 Sprague-Dawley rats into three groups(n=6):the control group(no special treatment),group OW(forced swimming twice a day for 15 days),and sleep deficiency(SD)+OW group(in addition to forced swimming twice a day,the rats were put on the platforms in water to limit sleep for 15 days).On the 16th day,the abdominal aorta and common carotid artery were collected after blood sampling from heart under deep anesthesia.A part of the abdominal aorta sample was taken for Masson staining of collagen fiber,and Verhoeff-Van Gieson staining was carried out for the elastic fiber of common carotid artery.Image J was employed for the quantitative analysis of collagen fiber and elastic fiber content.The expression of collagen 1(Col-1) protein was quantified by immunohistochemistry and the ultrastructure of vascular matrix was examined by transmission electron microscopy.The other part of the abdominal aorta sample was used to determine the mRNA levels of matrix metalloproteinase(MMP)-1,MMP-2,MMP-9,tissue inhibitor of metalloproteinases-1(TIMP-1),and Col-1 by quantitative real-time polymerase chain reaction. Results Compared with that in control group,the content of collagen fiber in groups OW and SD+OW had no significant change(all P>0.05);the content of elastic fiber in groups OW and SD+OW decreased(all P<0.001) and had no significant difference between each other(P>0.05).The vascular vessel wall of group OW showed slight fiber breakage,while that of group SD+OW presented wormhole-like or spongy fiber fragmentation.The mRNA levels of MMP-1 and MMP-2 in groups OW and SD+OW had no significant difference between each other(P>0.05) but were higher than that in control group(all P<0.001).The mRNA levels of MMP-9 and TIMP-1 had no significant difference among the three groups(all P>0.05).Groups OW and SD+OW had lower mRNA level(all P<0.001) and protein level(all P<0.001) of Col-1 than control group,while the mRNA and protein levels of Col-1 had no significant difference between groups OW and SD+OW(P>0.05). Conclusion OW can reduce the content of Col-1 and elastic fibers in the extracellular matrix of arterial vessels,destroy the elastic lamina of vascular wall,up-regulate the expression of MMP-1 and MMP-2,thereby injuring arterial vessels.

Efficacy of 25 gauge minimally invasive vitrectomy combined with cataract extraction and trabeculectomy surgeries for malignant glaucoma / 中华实验眼科杂志

Objective@#To evaluate the efficacy and outcome of 25 gouge (25G) minimally invasive vitrectomy combined with cataract extraction and trabeculectomy surgeries for malignant glaucoma.@*Methods@#Retrospective cohort study was performed.Clinical data of 19 malignant glaucoma patients (19 eyes) who received 25G minimally vitrectomy from January 2012 to January 2017 in Wuxi People's Hospital were reviewed retrospectively.The operative methods were selected according to the predisposing cause.25G vitrectomy combined with cataract extraction and posterior capsulotomy were performed on the malignant glaucoma eyes after trabeculectomy, and 25G vitrectomy combined with cataract extraction, trabeculectomy and posterior capsulotomy were performed on the malignant glaucoma eyes after non-trabeculectomy.Best corrected visual acuity (BCVA) was examined by international visual acuity chart.The ocular axis length and intraocular pressure (IOP) were measured with A-mode ultrasonic apparatus and non-contact tonometer, respectively.The anterior chamber depth was measured with ultrasound biomicroscope (UBM). The study followed the declaration of Helsinki and all patients signed informed consent before surgery.@*Results@#The operation was successfully completed in 19 patients.All patients suffered moderate to severe anterior chamber inflammation after operation.The average age of onset in the patients was (58.00±6.20) years, and the mean ocular axial length was (20.81±0.56)mm.Malignant glaucoma occurred in 11 eyes after trabeculectomy, 2 eyes after combination of anti-glaucoma with cataract extraction, 2 eyes after laser iridotomy, 2 eyes after paracentesis of anterior chamber and 2 eyes with unknown causes.The visual acuity was significantly improved 3 months after operation in comparison with before operation (Z=-3.826, P<0.001). The mean IOP was (12.16±2.27)mmHg (1 mmHg=0.133 kPa) in postoperation, which was significantly lower than (38.84±5.97)mmHg in preoperation (t=17.68, P<0.05). The depth of anterior chamber was increased from preoperative (0.95±0.28)mm to postoperative (2.43±0.15) mm (t=20.06, P<0.05). UBM image showed that the position of ciliary body was normal without edema.@*Conclusions@#The combination procedure of 25G minimally invasive vitrectomy with relative surgery for malignant glaucoma is effective by lowing IOP, rescuing visual acuity and reducing surgical risk.

The mechanism of repressive effects of transthyretitin on the growth of human retinal microvascular endothelial cells under high glucose and hypoxia environment / 中华眼底病杂志

Objective To explore repressive effects of transthyretitin (TTR) on the growth of human retinal endothelial cells (hREC) under high glucose and hypoxia environment. Methods hRECs were divided into 8 groups, including normal glucose group (5.5 mmol/L glucose), hypoxia group, high glucose group (25.0 mmol/L glucose), high glucose and hypoxia group, normal glucose group+TTR, normal glucose and hypoxia group+TTR, high glucose group+TTR, high glucose and hypoxia group+TTR. Flow cytometry was used to analyze cellular apoptosis. The expression level of Akt, p-Akt, eNOS, Bcl-2 and Bax protein were measured by Western blot. Results Hypoxia could induce apoptosis as the apoptosis rate of normal and hypoxia group was higher than normal group (χ2=25.360, P<0.05), high glucose and hypoxia group was higher that high glucose group (χ2=17.400, P<0.05). The cell apoptosis rate of high glucose and hypoxia group+TTR were increased significantly as compared with high glucose and hypoxia group (χ2=9.900, P<0.05). There was no statistically significant difference on the cell apoptosis rate between normal group and high glucose group, normal group+TTR and normal group, high glucose group+TTR and high glucose group, normal and hypoxia group+TTR and normal and hypoxia group (P>0.05). Western blot showed that the expression of Akt did not change significantly in all eight groups(F=2.450, P>0.05). Compared to normal group, the expression of p-Akt, eNOS, Bcl-2 in normal and hypoxia group were decreased (t=9.406, 5.306, 4.819), and the expression of Bax (t=-4.503) was increased (P<0.05). Compared to high glucose group, same trend was found in high glucose and hypoxia group (t=8.877, 7.723, 6.500, -14.646; P<0.05). The expression of p-Akt in normal and hypoxia group+TTR was higher than normal and hypoxia group (t=-5.024, P<0.05) ,but there was no difference on the expression of eNOS, Bcl-2, Bax between these two groups (t=-2.235, -2.656, -0.272;P>0.05). Compared to high glucose and hypoxia group, the expression of p-Akt and Bcl-2 in high glucose and hypoxia group+TTR were decreased (t=4.355, 4.308; P<0.05), the expression of Bax was increased (t=-4.311, P<0.05), and there was no difference on the expression of eNOS between these two groups (t=-1.590, P>0.05). There was no statistically significant difference in the expression of p-Akt, eNOS, Bcl-2, Bax between high glucose group and normal group (t=-3.407, -4.228, -4.302, -2.076; P>0.05), normal group+TTR and normal group (t=-4.245, -4.298, -2.816, -1.326; P>0.05), high glucose group+TTR and high glucose group (t=4.016, -0.784, 0.707, -0.328; P>0.05). Conclusion Under high glucose and hypoxia, transthyretitin suppress the growth of hREC through Akt/Bcl-2/Bax, but not Akt/eNOS signaling pathway.

Serum transthyretin in patients with different stages of diabetic retinopathy / 中华眼底病杂志

Objective To measure the concentration of serum transthyretin (TTR) of patients with different stages of diabetic retinopathy (DR).Methods A total of 176 patients with diabetes mellitus were included in this study.There were 104 males and 72 females.The patients aged from 21 to 74 years,with the mean age of(56± 11) years.The diabetes duration raged from 1 to 30 years,with the mean diabetes duration of (10 ± 7) years.The HbA 1C was 5.2％-14.1％,with the mean HbA 1C of (8.6 ± 2.0)％.According to the fundus examination,58 patients had DR (33.0％),but the other 118 patients not (67.0％).For these DR patients,10 patients were in stage Ⅰ (5.7％),26 patients in stage Ⅱ (14.8％),8 patients in stage Ⅲ (4.5％),and 14 patients in stage Ⅳ (8.0％).The concentration of serum TTR was measured by enzyme-linked immunosorbentassay kit.The differences in the concentration of serum TTR between different DR stages were compared.Bivariate analysis was used to analyze the influencing factors of TTR.Results The concentrations of serum TTR of the patients without DR or with DR of stage Ⅰ to Ⅳ were (224.96±65.47),(383.68± 102.99),(247.44±63.21),(228.2 ± 45.89),(189.34± 70.12) mg/L,respectively.The difference between different DR stages was statistically significant (F=14.690,P＜ 0.001).Bivariate analysis showed that the concentration of TTR was correlation to DR (r=0.179,P=0.017).There was no correlation between the concentration of TTR and diabetes duration (r=-0.027,P=0.727),hypertension (r=0.018,P=0.810),hyperlipoidemia (r=0.101,P=0.182),and the use of insulin (r=-0.032,P=0.675).Conclusion The concentration of serum TTR was increased in early DR patients,and gradually decreased with the progression of DR.The concentration of TTR is correlated to DR.

The mechanism of repressive effects of transthyretitin on the growth of human retinal microvascular endothelial cells under high glucose and hypoxia environment / 中华眼底病杂志

Objective To explore repressive effects of transthyretitin (TTR) on the growth of human retinal endothelial cells (hREC) under high glucose and hypoxia environment. Methods hRECs were divided into 8 groups, including normal glucose group (5.5 mmol/L glucose), hypoxia group, high glucose group (25.0 mmol/L glucose), high glucose and hypoxia group, normal glucose group+TTR, normal glucose and hypoxia group+TTR, high glucose group+TTR, high glucose and hypoxia group+TTR. Flow cytometry was used to analyze cellular apoptosis. The expression level of Akt, p-Akt, eNOS, Bcl-2 and Bax protein were measured by Western blot. Results Hypoxia could induce apoptosis as the apoptosis rate of normal and hypoxia group was higher than normal group (χ2=25.360, P<0.05), high glucose and hypoxia group was higher that high glucose group (χ2=17.400, P<0.05). The cell apoptosis rate of high glucose and hypoxia group+TTR were increased significantly as compared with high glucose and hypoxia group (χ2=9.900, P<0.05). There was no statistically significant difference on the cell apoptosis rate between normal group and high glucose group, normal group+TTR and normal group, high glucose group+TTR and high glucose group, normal and hypoxia group+TTR and normal and hypoxia group (P>0.05). Western blot showed that the expression of Akt did not change significantly in all eight groups(F=2.450, P>0.05). Compared to normal group, the expression of p-Akt, eNOS, Bcl-2 in normal and hypoxia group were decreased (t=9.406, 5.306, 4.819), and the expression of Bax (t=-4.503) was increased (P<0.05). Compared to high glucose group, same trend was found in high glucose and hypoxia group (t=8.877, 7.723, 6.500, -14.646; P<0.05). The expression of p-Akt in normal and hypoxia group+TTR was higher than normal and hypoxia group (t=-5.024, P<0.05) ,but there was no difference on the expression of eNOS, Bcl-2, Bax between these two groups (t=-2.235, -2.656, -0.272;P>0.05). Compared to high glucose and hypoxia group, the expression of p-Akt and Bcl-2 in high glucose and hypoxia group+TTR were decreased (t=4.355, 4.308; P<0.05), the expression of Bax was increased (t=-4.311, P<0.05), and there was no difference on the expression of eNOS between these two groups (t=-1.590, P>0.05). There was no statistically significant difference in the expression of p-Akt, eNOS, Bcl-2, Bax between high glucose group and normal group (t=-3.407, -4.228, -4.302, -2.076; P>0.05), normal group+TTR and normal group (t=-4.245, -4.298, -2.816, -1.326; P>0.05), high glucose group+TTR and high glucose group (t=4.016, -0.784, 0.707, -0.328; P>0.05). Conclusion Under high glucose and hypoxia, transthyretitin suppress the growth of hREC through Akt/Bcl-2/Bax, but not Akt/eNOS signaling pathway.

Comparison of axial length and anterior chamber depth obtained by A scan ultrasonography and IOL Master in different axial length groups / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To evaluate the repeatability of axial length (AL) and anterior chamber depth (ACD) obtained by A scan ultrasonography, and to compare AL and ACD obtained by A scan with those obtained by IOL Master.METHODS:Two hundred and fifty-seven cataract eyes of 170 patients were included.IOL Master and A scan were performed for each eye.Five measurements of IOL Master and 3 measurements of A scan were obtained.All the tested eyes were divided into 5 groups according to AL obtained by A scan:Group A (2129mm, 21 eyes).Cronbach's Alpha coefficient and intraclass correlation coefficient (ICC) were applied to evaluate the repeatability of AL and ACD obtained by A scan.Paired t test and Pearson correlation coefficient were used to analyze the differences and correlations of AL and ACD obtained by the 2 devices, respectively.Bland-Altman plots were presented to analyze the agreements of AL and ACD obtained by the 2 devices.RESULTS:All the Cronbach's Alpha and ICCs of AL and ACD values were more than 0.98.The differences of AL values between A scan and IOL Master were-0.11±0.08mm in Group A,-0.15±0.10mm in Group B,-0.19±0.15 mm in Group C,-0.29±0.16mm in Group D and-0.45±0.29mm in Group E, respectively (all P0.89, all P<0.01).CONCLUSION:The AL and ACD values in cataract eyes obtained by A scan were repeatable.The AL and ACD values obtained by A scan were smaller than those obtained by IOL Master.With the increase of AL values, the differences of AL values between A scan and IOL Master increased.

Inhibitory effects of apolipoprotein A-Ⅰ on biological behavior and VEGF expression of human retinal epithelial cells in high glucose environment / 中华实验眼科杂志

Background As a main cellular component of retinal microvascular,retinal vascular endothelial cells (RVECs) play critical roles in the occurrence and development of diabetic retinopathy (DR) by proliferating,migrating and angiogenesis.Apolipoprotein A-Ⅰ (ApoA-Ⅰ) is the major apolipoprotein of high density lipoprotein.ApoA-Ⅰ is overexpressed in the retina of diabetic patients and plays different effects on RVECs upon different microenvironments,but its relationship with RVECs in high glucose environment is still not elucidated.Objective This study was to investigate the effects of ApoA-Ⅰ on proliferation,migration,tubulogenesis and vascular endothelial growth factor (VEGF) expression in human RVECs (hRVECs) in high glucose environment.Methods hRVECs were cultured with DMEM containing 10％ fetal bovine serum and passaged,and the cells at generation 3 to 6 were used in the study.The cells were divided into low-glucose group,low-glucose+ApoA-Ⅰ group,high-glucose group and high-glucose+ApoA-Ⅰ group,and the low concentration glucose (5 mmol/L),high contration glucose (25 mmol/L)and ApoA-Ⅰ (30 μg/ml) was added separately according to grouping.The proliferation and migration rate of the cells were evaluated by cell counting kit-8 (CCK-8) assay and scratch wound test respectively.The tubulogenesis of the cells was examined by tube formation test.The mRNA and protein expression of VEGF in the cells was detected by using real-time fluorescence quantitative PCR and Western blot.Results The prolifative value (absorbancy) and migration rate of the cells in the high-glucose group were significantly higher than those in the low-glucose group,and those in the high-glucose+ApoA-Ⅰ group were significantly reduced in comparion with the high-glucose group (A value:P =0.001,0.033;migration rate:P =0.001,0.010).The number of tubes in the low-glucose group,low-glucose+ ApoA-Ⅰ group,high-glucose group and high-glucose+ApoA-Ⅰ group was 7.250±2.217,9.250±2.630,19.000± 3.916 and 11.500±3.697,showing a significant difference among the groups (F=10.335,P=0.001).The number of tubes in the high-glucose group was more than that in the low-glucose group,and the number of tubes in the highglucose+ApoA-Ⅰ group was less that that in the high-glucose group (P=0.001,0.037).The relative expression levels of VEGF mRNA were 0.944 ± 0.083,1.117 ± 0.204,1.768 ± 0.164 and 1.301 ± 0.077,and those of V EGF protein were 1.000±0.130,1.217±0.152,1.871 ±0.101 and 1.609±0.087 in the low-glucose group,low-glucose+ ApoA-Ⅰ group,high-glucose group and high-glucose+ApoA-Ⅰ group,respectively,with significant differences among the groups (mRNA:F =18.640,P =0.001;protein:F =10.335,P =0.001),and the expressions of VEGF mRNA and protein in the high-glucose group were significantly higher than those in the low-glucose group and high glucose+ ApoA-Ⅰ group (mRNA:P=0.000,0.004;protein:P=0.000,0.029).Conclusions ApoA-Ⅰ plays inhibitory effects on the proliferation,migration and tubulogenesis of hRVECs in high glucose environment,which may be associated with the downregulation of VEGF expression.