Objective To observe the effect of diabetic retinopathy on endothelial progenitor cells (EPCs)from peripheral blood.Methods Sixty male Wistar rats were divided into control group and diabetes group.The rats in diabetes group were induced with streptozotocin(STZ)injection for diabetic retinopathy model.Flow cytometry was used to identify and count the number of EPCs from peripheral blood at 1 week.1,3 and 6 months after injection.All eyeballs were examined by hematoxylin and eosin (HE)staining,periodic acid-Schiffs(PAS)staining of trypsin-digested retinal vessels flat preparation and transmission electron microscope.EPCs count,and the relationship between DR morphological changes and EPCs count were compared and analyzed.Results The quantity of EPCs from peripheral blood at 1 week,1,3 and 6 months after STZ injection were 25±7,28±8,39±7,43±7 cells per 200 000 monocytes respectively,which decreased compared with the control group 45±4 cells per 200 000 monocytes(F=8.933,P＜0.0 1).The quantity of EPCs was gradually increased at 1 week,1,3 and 6 months after STZ injection,accompanied with responsive pathological changes of retinal structure and vessels.The thickness of retina at 1 week and 1 month after injection were reduced slightly.The number of retinal ganglion cells reduced,with the time passing by.Endothelial cells were edema,mitochondrial was swollen,capillary basement membrane was thicken,lumen was significant stenosis,lumen occlusion and retinal artery aneurysm were observed at 6 months after STZ injection.Conclusion The number of EPCs increases gradually throughout the development of DR.

Different types of neurons are regularly dispersed on re tina. For finding the mechanism of eye and retina formation, it is important to know how t his regular pattern is formed during embryonic stage. It was claimed that multip le genes regulate the development of eye and retina. These genes regulate the de velopment of tissues and differentiation of cells in different parts of the visu al system during embryo development. Also, it was found that different tissues a nd differentiated cells cohere with each other both temporally and spatially to form the eye.

Objective To evaluate the effect of FTY720 on retinal leukocytes adhesion and vascular permeability in diabetic retinopathy (DR) model,and explore its mechanism.Methods Ninety male Wister rats were randomly divided into normal control group,diabetic group and FTY720 group,thirty rats in each group.Diabetes was induced by giving a single intraperitoneal injection of streptozocin.FTY720 group was administered with FTY720 at a dose of 0.3 mg/kg by oral gavage daily for 3 months after establishment of diabetes.All rats were used for experiments following intervention for 3 months in FTY720 group.Immunohistochemical staining was used to observe the expression and distribution of intercellular adhesion molecule (ICAM-1) and vascular cell adhesion molecule (VCAM-1),and the positive cells were counted.Real-time reverse transcription PCR was used to measure mRNA expression of ICAM-1 and VCAM-1.Fluorescein isothiocyanate-Concanavalin A perfusion was used to detect retinal leukocytes adhesion.Evans blue (EB) perfusion was used to analyze retinal vascular permeability.Immunofluorescence staining was used to detect retinal inflammatory cells infiltration.Results In diabetic group,both ICAM-1(t =12.81) and VCAM-1 (t=11.75) positive cells as well as their mRNA expression (t=16.14,9.59) were increased compared with normal control group,with statistical significance (P＜ 0.05).In FTY720 group,both ICAM-1(t=-9.93) and VCAM-1 (t=-6.61) positive cells as well as their mRNA expression (t=-15.28,-6.10) were decreased compared with diabetic group,with statistical significance (P＜ 0.05).Retinal leukocytes adhesion (t=16.32) and EB permeability (t=17.83) were increased in diabetic group compared with normal control group,while they were decreased in FTY720 group compared with diabetic group(t=-9.93,-11.82),with statistical significance (P＜0.05).There were many CD45 positive leukocytes infiltration in retina of diabetic group,including CD11b positive macrophage/activated microglia,while both of them were little in FTY720 group.Conclusions FTY720 can decrease retinal leukocytes adhesion,reduce retinal vascular permeability and inflammatory cells infiltration,which is associated with down-regulation of ICAM-1 and VCAM-1.

Objective To investigate the protective effect of Niacin on blood-retina barrier (BRB) in diabetic rats and related mechanism.Methods The male Wistar rats (60) were divided into control (CON) group,diabetes (DM) group and Niacin-treated (NA) group,20 rats in each group.Rats diabetes models were induced with streptozotocin injection.Niacin (40 mg/kg · d) was administrated orally everyday in Niacin-treated group until sacrificed after 3 months.Pathological outcomes,total cholesterol (TC) and highdensity lipoprotein (HDL) were evaluated at month 3.Optical microscopy was used to observe the retinal structure.The integrity of BRB and the vascular permeability was quantified by analyzing albumin leakage using Evans blue (EB) method.The relative expressions of Claudin-5,Occludin,zonula occluden (ZO)-1 and GPR109A mRNA in rat retinas were detected by reverse transcription PCR (RT-PCR) and relative expression of GPR109A,tumor necrosis factor (TNF)-α and interleukin (IL)-6 by Western blot.Results Compared to CON group,the TC content was increased and HDL content was decreased in DM group (t=4.034,5.831;P＜0.05).Compared to DM group,the TC content was decreased and HDL content was increased in NA group (t=6.868,3.369;P＜0.05).The retinal structure of CON group was normal.Pathological changes were found in the DM group,such as tumescent nuclei and disorganized structures.The retinal structure of NA group was similar to the control group.Evans blue dye that the microvascular leakage in DM group was increased compared with CON group (t=24.712,P＜0.05),while in NA group was decreased compared with DM group (t =16.414,P＜ 0.05).The mRNA expression of Occludin,Claudin-5,ZO-1 in DM group were decreased compared with CON group (t=11.422,12.638,12.060;P＜ 0.05),while in NA group were increased compared with DM group (t=5.278,3.952,8.030;P＜0.05).The mRNA expression of GPR109A in NA group were increased compared with DM group (t=5.053,P＜ 0.05).The protein expression of GPR109A,IL-6,TNF-αin DM group were increased compared with CON group (t=4.915,11.106,6.582;P＜0.05).Compared to DM group,the protein expression of GPR109A was increased (t=5.806,P＜0.05),while the protein expression of IL-6 and TNF-α were decreased (t=10.131,5.017;P＜0.05).Conclusion Niacin has the protective effect for BRB by up-regulating GPR109A expression which may suppress inflammation.

Objective To investigate the value of the levels of urinary NT-proBNP whether is similar with the levels of blood NT-proBNP for the diagnosis of chronic heart failure.Methods 79 patients with chronic heart failure (NYHA Ⅰ~Ⅳ)as a research group,and 53 healthy people as a control group,the levels of urinary and blood NT-proBNP on two groups were measured respectively by Roche cobas h 232 cardiac markers detector and compared the difference of the levels of urinary NT-proBNP and blood NT-proBNP between the two groups.Results The levels of urinary and blood NT-proBNP in pa-tients with chronic heart failure were much higher than those in healthy subjects,respectively (P <0.01).The concentration of urinary NT-proBNP increased gradually with more severe symptoms.The level of urinary NT-proBNP was positively cor-related with the levels of blood NT-proBNP.Conclusion The levels of urinary NT-proBNP is a new candidate marker for the diagnosis of chronic heart failures and it provides a similar accuracy with the levels of blood.

BACKGROUND:Matrix metal oproteinases and their inhibitors are proteolytic enzymes contaning Zn+, and involved in extracel ular matrix degradation and tissue remodeling of a variety of tissues. OBJECTIVE:To observe the effects of matrix metal oproteinases in the proliferation of MC3T3-E1 cel s induced by lipopolysaccharide. METHODS:MC3T3-E1 cel line was divided into four groups randomly:control group, low-dose lipopolysaccharide group (1μmol/L), moderate-dose lipopolysaccharide group (10μmol/L), and high-dose lipopolysaccharide group (100μmol/L). The proliferation rate in each group was analyzed. Matrix metal oproteinase 2, 3, 9 and matrix metal oproteinase inhibitor 1 and 2 expressions were detected. RESULTS AND CONCLUSION:The proliferation rate was increased greatly after medication of lipopolysaccharide in time-dependent and concentration-dependent manners. Moreover, the expressions of matrix metal oproteinases and their inhibitors were apparently enhanced, and showed significant differences. Results indicate that the enhanced expressions of matrix metal oproteinases participated in the proliferation of MC3T3-E1 cel s induced by lipopolysaccharide.

Objective To compare the effects of intravitreal tamponade of C3 F8 with silicon oil on postoperative vitreous hemorrhage and visual prognosis after vitrectomy for proliferative diabetic retinopathy (PDR).Methods The clinical data of 121 patients (127 eyes)who underwent primary vitrectomy due to PDR were analyzed retrospectively.All the patients were divided into two groups according to different intravitreal tamponade, including C3 F8 tamponade group (53 patients with 56 eyes ) and silicone oil tamponade group (68 patients with 71 eyes).There was no difference of gender (χ2 = 0.956 ),age (t =1.122),duratiion of diabetes (t=0.627),fasting blood glucose (t=1.049),systolic pressure (t=1.056), diastolic pressure (t = 0.5 1 7 ), history of hypertension (χ2 = 0.356 ), nephropathy (χ2 = 1.242 ), preoperative laser photocoagulation (χ2 = 1.225 )and All the patients underwent three port pars plana vitrectomy.The mean follow-up was 2 years ranging from 6 months to 4 years.And then the incidence and onset time of postoperative vitreous hemorrhage and postoperative BCVA of the two groups were compared. Results Postoperative vitreous hemorrhage occurred in 14 of 56 eyes (25.00%)in C3 F8 tamponade group. The average onset time of postoperative vitreous hemorrhage were (64.64 ± 59.09)days ranging from 7 -225 days and mostly were within 30-60 days (35.71%,5/14).Postoperative vitreous hemorrhage also occurred in 7 of 71 eyes (9.89%)of silicone oil tamponade group after silicone oil removal with an average onset time of (25.29±20.46)days ranging from 3-65 days and were mostly within 1 5-30 days (42.86%, 3/7).There was a significant difference in the incidence of postoperative vitreous hemorrhage between the two groups (χ2 = 5.200,P <0.05 ).BCVA of the two groups was improved significantly after operation (Z =2.472,3.1 14;P <0.05).Postoperative BCVA of silicone oil tamponade group was poorer than C3 F8 tamponade group (Z =1.968,P <0.05).Conclusion Both C3 F8 and silicone oil tamponade can improve the visual acuity after vitrectomy for PDR.Compared with C3 F8 ,silicone oil tamponade had lower incidence and late onset of postoperative vitreous hemorrhage after vitrectomy for PDR.

Background The signal pathway of mammalian target of rapamycin (mTOR) plays an important role in the regulation of cell cycle.Rapamycin(RAPA) can inhibit the proliferation of cells by regulating of cell cycle.Objective This study was to investigate the effect of RAPA on the proliferation of Rhesus retinal vascular endothelial cells (RF/6A).Methods RF/6A cell strains were cultured in vitro,and PBS,10 μg/L RAPA or 5 μg/L RAPA was added into the medium respectively.The expression of cyclin D1 protein in the RF/6A cells (absorbancy) were detected by Western blot.The cell cycle distribution after RAPA action was analyzed by flow cytometry.Matrigel was used in endothelial-cell tube formation to evaluate the effect of RAPA on angiogenesis.Results Western blot assay showed that the expressions of cyclin D z protein were 0.92±0.04,0.58±0.02 and 0.73±0.02 in the PBS group,10 μg/L RAPA group and 5 μg/L RAPA group,showing a significant difference among the three groups (F =246.320,P =0.000),and the relative expressing level of cyclin D1 protein in the l0 μg/L RAPA group and 5 μg/L RAPA group was significantly lower than that of the PBS group (both at P＜0.05).The proportion of G0/G1 cells were (42.13±0.57)％,(65.15±0.64)％ and (54.09± 0.78)％ respectively in the PBS group,10 μg/L RAPA group and 5 μg/L RAPA group,which was significantly different among the three groups (F=887.815,P=0.000).The number of endothelial-cell tubes were (9.67 ± 1.53)/field,(4.33 ± 0.58)/field and (6.33 ±0.58)/field respectively in the PBS group,10 μg/L RAPA group and 5 μg/L RAPA group,and the number of endothelial-cell tubes in 10 μg/L RAPA group and 5 μg/L RAPA group was less than that in the PBS group,with a significant difference among the three groups (F =21.778,P =0.002).Conclusions RAPA can inhibit the proliferation of RF/6A cells in vitro by down-regulating the expression of cyclin D1.

Background Chrysin has many biological activities,and its anti-inflammatory effect has been confirmed.However,whether it can treat experimental autoimmune uveitis (EAU) is still not elucidated.Objective This study was to investigate the therapeutic effect of chrysin on EAU and explore the potential mechanism.Methods EAU animal models were established in 30 SPF C57BL/6J mice by the subcutaneous injection and ball pad injection of interphotoreceptor retinoid binding protein1-20 (IRBP1-20)/complete Freund adjuvant (CFA),and then the models were randomized into the model control group and chrysin-treated group.Chrysin solution dissolved by 10 μl dimethyl sulfoxide (DMSO)+140 μl PBS was administrated by gavaging in the mice with the dlose 25 mg/kg from 3 days before immunization through 21 days everyday in the chrysin group,and equal volume of solvent was used in the same way in the model control group.Retinas were examined by indirect ophthalmoscope once per day,and inflammation and pathological scores of retina were performed based on the criteria of Caspi on the 21st day after injection.The apoptosis of retinal cells was assayed by TUNEL staining,and the relative expressions of proinflammatory cytokines interferon-γ (IFN-γ),interleukin-17 A (IL-17A),IL-6 and tumor necrosis factor-α (TNF-α) and signal transducer and activator of transcription 1 (STAT1),STAT3,p-STAT1,p-STAT3 in mouse retinas were detected by Western blot.Results Compared with the model control group,the inflammation scores and pathological scores of retinal inflammation were significantly reduced in the chrysin group (inflammation scores:0.50± 0.45 vs 1.58±0.92,t=2.600,P=0.026;pathologic scores:0.58±0.38 vs 1.83±0.75,t=3.638,P=0.005).The infiltration of a large number of inflammatory cells,retinal vasculitis and granulomatous lesions were found in mouse retinas in the model control group,however,the morphology of mouse retinas in the chrysin group was normal based on hematoxylin-eosin staining.The number of apoptotic cells was remarkable lessened in the chrysin group compared with the model control group under the fluorescence microscope.Western blot assay resolved that significantly downregulation in the expressions of IFN-γ,IL-17A,IL-6 and TNF-α was seen in the chrysin group in comparison with the model control group (t =7.802,P =0.001;t =14.906,P =0.000;t =10.241,P =0.001;t =3.304,P =0.030),and the relative expression levels of STAT1,STAT3,p-STAT1 and p-STAT3 were considerably lower in the chrysin group than those in the model control group (t=8.965,P=0.001;t=8.358,P=0.001;t=4.864,P=0.031;t=4.730,P=0.009).ConclusionsChrysin or chrysin-like flavonoids ameliorate intraocular inflammatory symptoms in EAU mice by inhibiting the activity of STAT signal pathway.

Background Studies showed that traumatic optic neuropathy (TON) can activate and induce immuno-inflammatory response,while T helper cell 17 (Th17), a CD4+ T lymphocyte, is associated with immunoinflammatory response.However,the effects of Th17 on the development of TON are unclear.Objective This study was to observe the changes of Th17 lymphocytes in spleen and the expression of interleukin-17 (IL-7) in retina in TON rats.Methods Seventy male 4-week-old SD rats were randomly divided into normal control group, sham operation group and TON 1-, 3-,7-, 14-and 28-day group according to random number table.The optical nerves of the right eyes were exposed and impacted at a 25° angle by using fluid percussion injury (FPI) device with the average force (699.1±60.8) kPa to create the TON models in the TON groups,and only optical nerves were exposed in the sham operation group.Flash visual evoked potential (F-VEP) was recorded and the amplitude and latency were measured to evaluate the function of optical nerves of the rats.The rat splenocyte suspension in various groups was prepared for the detection of Th17 cells by flow cytometry, and the expression of IL-17 protein was detected by immunochemistry.Results No significant differences were found in the amplitudes and latencies of P2 wave, proportion of Th17 cells and the IL-17 positive cell numbers in retina between the normal control group and sham operation group (P =0.829,0.830,0.856,0.496).The amplitudes of P2 wave were significantly lower, and the latencies of P2 wave were significantly longer in the TON groups than those in the sham operation group (all at P＜ 0.05).The proportions of Th17 positive CD4+ T lymphocytes were (0.94±0.13)％,(1.80±0.18)％,(1.98± 0.20)％ ,(2.34±0.20)％ ,(2.11±0.13)％ ,(1.92±0.18)％ in the sham operation group,TON 1-day group,TON 3-day group,TON 7-day group,TON 14-day group and TON 28-day group, respectively, showing significant differences between the sham operation group and various TON groups (all at P＜0.05).In addition, the IL-17 positive cell numbers in retina were 796.326 ± 100.028, 1 673.416 ± 188.021,1 892.431 ± 151.026,2 420.454 ± 256.024,1 996.429±177.022 and 1 629.410±127.023 in the sham operation group,TON 1-day group,TON 3-day group,TON 7-day group,TON 14-day group and TON 28-day group, respectively, showing significant differences between the sham operation group and various TON groups (all at P＜0.05).Conclusions In rats of TON, the proportion of Th17 cell in splenocytes and the expressions of IL-17 in retina are increased,suggesting that Th17 and IL-17 participate in the progression of TON.