Background 23G pars plana vitrectomy has been widely applied to treat diabetic retinopathy (OR).Researching the influence of 23G pars plana vitrectomy on corneal endothelium cell has a great clinical significance.Objective To observe the influence of 23G pars plana vitrectomy on corneal endothelial cells in phakic eyes of diabetes and non-diabetes patients.Methods A retrospective study was designed.One hundred and twenty-four eyes of 124 patients with vitreoretinopathy were included in Affiliated Hospital of Qingdao University from August 2012 to June 2013.The patients were assigned to DR group (52 eyes) and non-DR group (72 eyes).23G pars plana vitrectomy was performed on all the patients under their informed consent.Endothelial cell density,corneal thickness(CT),coefficient of variation (CV) of cellular area,standard deviation (SD) of average cellular area and percentage of hexagonal endothelial cells were measured before and 1 day,3 days,1 week,2 week,1 month and 3 months after surgery with Topcon SP-3000P corneal specular microscope.Results No significant differences were found in the central corneal endothelial cell density between the DR group and non-DR group at various time points (Fgroup =2.148,P=0.150;Ftime =0.900,P=0.504).The CV of endothelial cells,SD of endothelial cellular area and CT in the first day after surgery were higher than preoperation (P =0.000,0.011,0.033),while the percentage of hexagonal endothelial cells was declined (P =0.001).The CV of endothelial cells and the percentage of hexagonal endothelial cells recovered in postoperative 1 month in the DR group.In the non-DR group,the CV of endothelial cells and CT elevated in postoperative 1 day in comparison with preoperation (P =0.002,0.003),and the percentage of hexagonal endothelial cells reduced (P =0.000).These abnormalities returned to a preoperative level in a week after surgery.Conclusions 23G pars plana vitrectomy results in a reversible morphology damage of corneal endothelial cells.These damage may be more severe with a longer duration in DR patients compared with non-DR patients.

Objective To observe the changes of retinal morphology and function of macular-off rhegmatogenous retinal detachment (RRD) after scleral bulking.Methods In this prospective study,42 eyes of 41 patients who underwent scleral bulking were enrolled.There were 26 males (27 eyes) and 15 females (15 eyes),with an average age of (33.78± 11.21) years.Best corrected visual acuity (BCVA),intraocular pressure,indirect ophthalmoscope,visual fields,optical coherence tomography (OCT) and B scan of ocular ultrasound were measured for all patients.The average BCVA was 0.29±0.18.The retinal detachment time was (21.12±3.71) days.The mean visual field defect (MD) was (13.54±6.44) dB.The mean loss variance (LV) was (8.43±2.11) dB.All the patients were performed cryotherapy and sub-choroidal fluid drain out.The mean follow up was 12.4 months (from 6 to 24 months).At two weeks,1,3,6,12 months after surgery,the changes of BCVA,visual fields,retinal morphology and subretinal fluid were observed.Results Indirect ophthalmoscope combined with B scan showed the time of retinal reattachment was (7.32±2.53) days.Subretinal fluid was found completely absorbed by OCT with a mean of (7.82±3.52) months.At 12 months after surgery,subretinal fluid was completely absorbed in 37 eyes (88.10％).In these 37 eyes,15 eyes had normal retinal microstructure,5 eyes had neuroepithelial cystoid edema; 12 eyes had disrupted inner segment/outer segment (IS/OS) junction,and 5 eyes had disrupted IS/ OS and external limiting membrane (ELM).BCVA at 6 months after surgery was no significant difference with that at 12 months after surgery (t=-0.636,P=0.529).At 12 months after surgery,there were4 retinal patterns on OCT examination,including normal retinal microstructure,neuroepithelial cystoid edema,IS/OS line disruption,and IS/OS and ELM disruption.The BCVA difference among these 4 groups was significant (F =52.42,P ＜ 0.05).The BCVA difference between eyes with or without residual subretinal fluid was significant (t=-5.747,P=0.000).At 1,2 weeks and 1,3,6,12 months after surgery,the MDwere (11.38±2.53),(10.14±2.19),(9.17±2.13),(6.63±1.70),(5.71±1.89),(5.14± 1.69) dB respectively,with a significant difference between these time-points (F=63.528,P =0.00).However,the MD at 6 months after surgery was no significant difference with that at 12 months after surgery (t=1.442,P=0.157).At 12 months after surgery,there were 12 eyes with normal MD,30eyes with higher MD.There was no significant difference between surgery eyes with higher MD and fellow eyes in MD (t =-1.936,P =0.06).The MD value was positively correlated to the time of retinal detachment in patients with normal retinal microstructure (r=0.84,P =0.00).There were differences in LV during different periods after surgery (F=57.25,P =0.00).Conclusions The retinal microstructure,visual acuity,visual fields were gradually improved after scleral bulking.The patients had better vision with normal retinal microstructure.The time of retinal detachment positively correlated with visual fields damage.

OBJECTIVE:To improve HPLC for content determination of ketoprofen in Ketoprofen enteric-coated capsules. METHODS:HPLC method was adopted. The determination was performed on Chiralpak IC column with mobile phase consisted of n-hexane (0.1% TFA)-isopropanol(90:10,V/V)at a flow rate of 0.8 mL/min. The detection wavelength was set at 268 nm,and column temperature was 25 ℃. The sample size was 10 μL. RESULTS:The linear range of ketoprofen were 0.025-0.5 mg/mL(r=0.9998). The limit of quantitation was 1.0 mg/L,and limit of detection was 0.2 mg/L. RSDs of precision,stability and reproduc-ibility tests were lower than 2%;recoveries were 96.36%-100.32%(RSD=1.87%,n=6). CONCLUSIONS:The method is sim-ple,accurate and rapid,and can be used for the content determination of ketoprofen in Ketoprofen enteric-coated capsules.

Objective o observe the expression of Notch1 and Delta-like ligand 4 (Dll4) on the fibrovascular membranes in proliferative diabetic retinopathy (PDR),and investigate its relationship with vascular endothelial growth factor receptor 2 (VEGFR2).Methods Fifty-seven PDR patients (60 eyes) who underwent vitrectomy were enrolled in this study.The PDR patients were divided into non-injection group (30 patients,32 eyes) and injection group (27 patients,28 eyes).The eyes in injection group received intravitreal injection with ranibizumab at 2 to 7 days before surgery.The preretinal fibrovascular membranes were obtained from the PDR patients during vitrectomy.Eighteen epiretinal membranes were obtained from the non-diabetic patients was served as controls.The real-time polymerase chain reaction (RT-PCR) and immunohistochemical methods were used to detecting the expression ofNotch1,Dll4 and VEGFR2.In the meantime,the numbers of the nucleus of vascular endothelial cells in the membranes stained with hematoxylin were counted.Results The immunohistochemical staining revealed that there were positive expression ofNotch1,Dll4 and VEGFR2 in all PDR membranes,regardless of the injection of the ranibizumab.The levels ofNotch1,Dll4 and VEGFR2 protein in non-injection group were higher than those of injection group (t=3.45,6.01,4.08;P=0.030,0.008,0.023).In injection group,the number of endothelial cells in the membranes reduced (17.17 ± 2.48) compared with that of the non-injection group (41.50± 5.57).There was significant difference in the number of endothelial cells in the membranes between the two groups (t=9.58,P＜0.05).RT-PCR showed that the differences of the mRNA expression ofNotch1,Dll4 and VEGFR2 were all statistically significant among the PDR group and control group (H=12.50,12.50,12.02;P＜0.05).The expression ofNotch1,Dll4 and VEGFR2 in the PDR membranes was higher than that of epiretinal membranes from non-diabetic patients.In the PDR group,the expression of Notch1,Dll4 and VEGFR2 of non-injection group was higher than that of injection group.Spearman correlation analysis showed that the expression of mRNA between VEGFR2 and Dll4 (r=0.83),VEGFR2 and Notch1 (r=0.81),Notch1 and Dll4 (r=0.87) were all significantly correlated (P＜0.05).Conclusions The expression of Notch1 and Dll4 in the PDR membranes are higher than that of the control group,and it is positively correlated with the expression of the VEGFR2.Notchl and Dll4 play a regulatory rule in the neovascularization in PDR,the acting way may be correlated with VEGFR2.

Objective To observe the effect of different concentration netrin-1 on retinal vascular permeability in diabetes mellitus (DM) rats.Methods Eighty adult Sprague-Dawley rats were randomly divided into 8 groups,10 rats in each group,including normal control group (group A),normal+balanced salt solution (BSS) group (group B),normal+netrin-1 (500 μg/ml) group (group C) and DM group (50 rats in 5 subgroups).DM rats were induced by intraperitoneal injection of streptozocin.Three months after intraperitoneal injection,10 DM rats in the control group were injected with BSS (group D).Forty DM rats were injected with 5 μl of different concentrate netrin-1,and were divided into DM+netrin-1 10 μg/ml group (group E),DM+netrin-1 50 μg/ml group (group F),DM+netrin-1 100 μg/ml group (group G),DM+netrin-1 500 μg/ml group (group H)according to the different concentration.Non-DM rats in group C were injected with netrin-1 500 μg/ml.The expression of occludin was determined by immunohistochemistry for protein,and by real-time fluorescence quantitative reverse transcription polymerase chain reaction for mRNA level.Retinal vascular permeability was measured by Evans blue infusion.Results The expression of occludin protein and mRNA in group D were less than group A (t=27.71,8.59;P=0.00,0.00).However,the retinal vascular permeability increased in group D (t=-42.72,P=0.00).The expression of occluding protein,occludin mRNA and retinal vascular permeability showed significant differences between group D,E,F,G and H (F=146.31,16.54,67.77;P=0.00,0.00,0.00).Compared the group B with group C,there was no significant differences between the expression of occludin protein,occludin mRNA and the retinal vascular permeability (t=-1.13,0.93,1.04;P=0.27,0.36,0.31).The concentrate of netrin-1 showed a significant positive correlation to the expression level of occludin and occludin mRNA (r=0.73,0.81;P=0.00,0.00),but negative correlation to the vascular permeability (r=-0.61,P=0.00).Conclusion Netrin-1 can reduce the DM rats' retinal vascular permeability,which depended on the concentration of netrin-1.

OBJECTIVE To make a new sterilizer to disinfect the instruments used for the root pulp treatment.METHODS After studying the SL sterilizer used in Japan,a new sterilizer was made.Studied how long the sterilizer could be used after putting through the electric.The effect of it was compared with the Eurouda sterilizer E6-18/24 by the method of culturing the bacteria.RESULTS 208.8 seconds after putting through the electric,the sterilizer could be used to sterilize the apparatus.And the effect of the new sterilizer was the same as the Eurouda sterilizer E6-18/24.CONCLUSIONS The effect of the sterilizer is testified,and the sterilizer could be used for the root pulp treatment instruments.

The expression of paired immunoglobulin-like receptor B (PirB) in normal and injured spinal cord of rats was investigated. The SD rat hemi-sectioned spinal cord injury (SCI) model was established. Before and 1, 3, 7, 10 days after SCI, the spinal cord tissues were harvested, and Western blot and immunohistochemistry were used to examine the expression and location of PirB. The results showed that the expression level of PirB in the normal spinal cord of SD rats was low. At the first day after SCI, the expression of PirB was obviously increased, and that in the injured spinal cord from the first day to the 10th day was significantly higher than in the normal spinal cord. The positive expression of PirB in neurons from different regions of gray matter of the injured spinal cord was seen. It was concluded that the expression of PirB in the normal spinal cord of rats was low. The expression of PirB in SCI was significantly increased till at least the 10th day.

[Objective] To investigate the mesenchymal stem cells (MSC) in treating acute lung injury (ALI) via ALI mouse model.[Methods] By monoclonal antibody Anti-GD2 of specific antigen ganglioside (GD2) only expressed on MSC as a carrier,new fluorescent molecule probe were synthesized through covalently coupling Anti-GD2 and fluorescent group CyDye mono-reactive NHS Esters (Cy7).Synthetic Anti-GD2-Cy7 and MSC were labeled by the specific binding of antigen and antibody in vitro.Total 84 balb/c male mice were selected and randomly selected 48 mice were divided into three groups:sham group (n =16),MSC+ ALI group (n =16),NS + ALI group (n =16).The lung histopathology and scores,lung W/D ratio and permeability of lung microvasculature were examined at 24 h,48 h after ALI mouse model.Other 36 mice were randomly divided into three groups:normal group (n =12),sham group(n =12),MSC +ALI group(n =12).Labeled MSC-GD2-Cy7 were transplanted into MSC+ALI group and sham group mice via tail vein injection.At 30 min,1 d,3 d,and 7 d post-MSC-GD2-Cy7 injection,the mice were sacrificed after anesthesia and then the lung was removed.Excised lung was detected on small animal fluorescent imager.[Results] Contrast to NS+ ALI group,the lung histopathology and scores,lung W/D ratio and permeability of lung microvasculature of MSC +ALI group were more greatly improved at both 24 h and 48 h.Fluorescent results showed that the signal intensity in thc lung of MSC +ALI group was significantly higher than that of sham group at each time point [(3.37 ± 0.02)× 10-4 vs (2.05 ± 0.04) × 10-4 scaled counts/s;(35.54 ± 0.47)× 10-4 vs (25.29 ± 1.48) × 10-4 scaled counts/s;(11.17 ± 0.75)×10-4 vs (6.09 ± 0.62)× 10-4 scaled counts/s;(3.10 ± 0.14) vs (0.00 ± 0.00)× 10-4 scaled counts/s;all P ＜ 0.05].[Conclusion] Our study showed that a proportion of cells migrated into normal and injured lungs 30 min after cell transplantation,and the cells started to recruit and largely gather in injured lungs at day 1 and persisted to day 7,these results suggest that MSC possess the ability to home into injured tissues.

Objective To investigate the methylation status of E-cadherin(E-cad), p16, RASSF1A, DAPK and MGMT in histologically normal salivary gland tissues and provide reference for determination of the methylation status of salivary gland tumors.Methods Methylation of E-cad, p16, RASSF1A,DAPK and MGMT was analyzed using methylation-specific polymerase chain reaction ( MSP) .The results were compared with the methylation status of these genes in salivary adenoid cystic carcinoma ( ACC) tumor tissues in our previous studies and the association between promoter methylation of E-cad, p16, RASSF1A, DAPK, and MGMT on one hand and the patients′gender, age, smoking and types of gland on the other hand was also analyzed .Results Promoter methylation was detected in 8 of the 60 (13%) salivary glands, E-cad in 4(7%), p16 in 2(4%), RASSF1A in 2(4%), DAPK in 2 (4%), and MGMT in 1(2%).Compared with our previous results, there was a significantly lower methylation ratio in promoter methylation of E-cad(P<0.01), p16 (P<0.01), RASSF1A (P<0.01),and DAPK (P<0.01) in salivary gland tissues than in ACC tumor tissues.Conclusion Promoter methylation of E-cad, p16 and RASSF1A is a rare event in histologically normal salivary gland tissues .

Epigenetics palys a important role in many dieases,including diabetic retinopathy (DR).Epigenetic modifications include DNA methylation,histone modifications and deployment of noncoding RNA,which are related to DR.Elucidateing the epigenetic mechanism of DR may provide a new direction for treating diabetes.