Objective To investigate the change of serum level of IL-6 and TNF-? in patients with Graves disease(GD). Methods Serum IL-6 and TNF-? levels of 37 GD patients before and after treatment were measured. And 30 healthy subjects served as controls. Results Serum levels of IL-6 and TNF-? in GD patients before treatment were significantly higher than those in the controls(P

Purpose: A comprehensive quality assurance program has been established in Beijing hospital to ensure that "radiosurgery" be carried out precisely and safetely.Materials and Methods: A film checking technique was used to verify the localization accuracy and the setting-up accuracy.Results: The figures taken from 80 cases treated show that the setting-up accuracy of the target positions be within ?1mm .Conclusion: The positional accura cies during target localization and setting-up are guaranteed by using our QA procedures.

Objective To establish a matrine delivery system in vitreous is very important for the dynamic treatment of proliferative vitreoretinopathy(PVR) . Present study was to evaluate the efficacy of matrine polyactic acid microsphere(MAT-PLA-MS) in prevention of PVR. Methods The suspension of cultured fibroblasts was injected into vitreous cavity of 30 healthy adult New Zealand albino rabbits to induce PVR. Then the experimental rabbits were divided into 3 groups and 10 rabbits for each. The animals received intravitreal injection of 0.3 mL MAT-PLA-MS(4 mg) matrine in MAT-PLA-MS group. Free matrine normal sodium solution 0.3 mL(containing 2mg matrine) was injected in vitreous cavity in free matrine group. 0. 3 mL normal saline solution was injected into the vitreous of the left eyes and the equivalent volume of blank polyaetic acid microsphere(blank-PLA-MS) into the right eyes in control group. The changes of cornea, aqueous humor, lens, vitreous and fundus were examined and recorded by slit lamp biomicroscope, indirect ophthalmoscope, fundus color camera and B ultrasonogram on the 1st, 3rd, 7th, 14th, 21st, 28th and 35th day following injection of drug. The inhibition effect of matrine on PVR was evaluated according to Ryan' s grading criteria of PVR. Results On the 14th days after implantation of MAT-PLA-MS, the rate of retinal detachment was 60%, 10%, 5% and 60% in normal saline group, free matrine group, MAT-PLA-MS group and blank-PLA-MS group respectively. Statistically significant difference was found among normal saline group, blank-PLA-MS group, MAT-PLA-MS group and free matrine group(P ＜0. 05). On the 21st day after injection of fibroblasts, the morbidity of retinal detachment was 80%, 30%, 10% and 80% in normal saline group, free matrine group, MAT-PLA-MS group and blank-PLA-MS group respectively, showing a significant difference among different groups. On the 28th day, the incidence rate of retinal detachment was 90%, 50%, 15% and 90% respectively, presenting statistical difference among various groups (P ＜ 0. 05) as well as between free matrine group and MAT-PLA-MS group (P＜0. 05). On the 35th day, considerably difference also was seen in the morbidity of retinal detachment among various groups (90%, 60%, 15% and 90% respectively) (P＜0.05). Conclusion Implantation of MAT-PLA-M S into vitreous cavity can effectively inhibit the development of PVR induced by fibroblasts in rabbit model.

Background　Culture of retinal pigment epithelium(RPE) cells is very important for establishment of proliferative vitreoretinopathy (PVR) model,prevention and treatment of PVR as well as RPE cell transplantation.Isolation of animal RPE cells by trypsinization is a critical step.ObjectiveThe present study is to establish the methods of isolation and culture of retinal pigment epithelium (RPE) cells in rabbit and comparied with that of pig RPE culture.MethodsRPE cells were isolated by trypsinization in pigmented rabbit and pig and cultured in DMEM containing 20% fetal bovine serum.Cultured RPE cells were identified by immunochemistry.The fourth generations of cells were used in this experiment.Morphology and characteristics of cultured RPE cells from rabbit and pig were examined and compared under the light microscope.ResultsIsolated RPE cells from pig were obtained by once trypsin digestion,but two times of trypsinization were needed in rabbit RPE cells isolation.The differentiation in response to trypsinization was related to anatomic difference between the two types of cells .The adherence time of pig RPE cells was 24 hours ,however,the rabbit RPE needed 48-72 hours after culture.Proliferation and vitality of cultured cells were gradually attenuated and melanin decreased after several times subculture.The morphology of culture RPE cells was obviously different between rabbit and pig because species difference.Immunohistochemistry demonstrated the positive response of RPE cells for keratin.ConclusionRPE cells can be acquired from both rabbit and pig by trypsinization and culture.The culture process of RPE cells of pig is simpler than that of rabbit.Cells within the fourth generations are suitable for experimental application.

Objective To investigate the effect of different doses of protamine neutralizing heparin on perioperation of on-pump coronary artery bypass graftting (CABG).Methods A total of 180 on-pump CABG patients hospitalized from January 2015 to November 2016 were randomly divided into three groups,the protamine group l,protamine group 2 and protamine group 3,60 patients in each group.Heparin (3 mg/kg) was used before extracorporeal circulation.After intracardiac operation was over,protamine was used to neutralize the heparin to adjust the activated clotting time (ACT) in protamine group 1,which was 10％ higher than that of intubation.Meanwhile,protamine group 2 was neutralized to equal to the ACT before intubation,and protamine group 3 was 10％ lower than that before the intubation.The differences of intraoperative and postoperative parameters were compared between the three groups.Results No death was found in the three groups during hospitalization.Comparing with protamine group 1 and protamine group 2,the time of operation,the ACT before the leaving operation room,the ACT of the first hour after returning to ICU,the amount of bleeding during operation,the time of closing and the amount of red blood for transfusion were decreased in protamine group 3 (P ＞ 0.05).The total amount of protamine for neutralizing and the ratio of protamine and heparin were significantly increased in protamine group 3 (P ＜ 0.05).The heart dysfunction after operation,perioperative myocardial infarction,pulmonary edema,pulmonary infection,renal dysfunction,poor wound healing,neurological complications,and time of in hospital stay showed no significant differences between three groups (P＞0.05).Conclusion ACT below 10％ of preoperation is safe,after neutralization of heparin by protamine,which can obviously reduce the bleeding,the time of sternal closure and the amount of red blood cell transfusion,showing a positive clinical significance.

Background The anti-proliferative effect of matrine has been demonstrated and its relevance to prevention and treatment of proliferative retinovitreopathy is concerned.Howeverthe intravitreous injection of free-matrine reiteratively may raise the risk of ocular infection.ObjectiveThe goal of the present study is to investigate the sustained releasing ability and safety of matrine polyactic acid microsphere(MAT-PLA-MSintravitreal injection.MethodsMAT-PLA-MS was prepared by Hebei Medical University and examined under the transmission electron microscope.The release of MAT-PLA-MS was monitored by ultraviolet spectrophotometry.Free-matrine with the dose of 1,2,4mg was intravitreally injected respectively in 12 eyes of New Zealand albino rabbits in free-matrine group and MAT-PLA-MS with matrine(2,4,6mg respectively was administered in 16 eyes separately in matrine microsphere group.The blank microsphere was injected in 6 right eyes as blank control group and normal saline solution was injected in 6 fellow eyes as control group.The retinal function change was evaluated by electroretinogram(ERG),and the morphological and histological change of retina following drug injection were assessed under the slit lamp biomicroscope,indirect ophthalmoscope,light microscope and transmission electron microscope.The decomposed process of MAT-PLA-MS in vitreous was recorded with ocular anterior segment and fundus color camera.Results MAT-PLA-MS containing matrine showed the spherical shape with the mean diameter of 2.28±47μm under the transmission electron microscope and the drug-loading rate 6.17% and drug-release rate 87.93% in vitro for 672 hours,presenting the controllable release characteristics.After implantation into the vitreous,the MAT-PLA-MS containing matrine decomposed gradually with the prolong of time.The b amplitudes of ERG maximum response were significantly declined in 4mg free-matrine injection group in comparison with before injection in various time points(P<0.01).However,no considerably differences were found in MAT-PLA-MS with matrine groups and control groups in various time points following the intravitreal injection(P>0.05).No obvious abnormal was seen under the slim lamp and ophthalmoscope through the study period.The changes of retinal ultrastructure were found from 1 through 28 days after injection of 4mg free-matrine,and slight retinal structural damage was seen from 7 through 28 days after injection of 6mg MAT-PLA-MS containing matrine.ConclusionThese results suggest that MAT-PLA-MS possesses good sustained release feature.MAT-PLA-MS containing matrine has less toxicity to retina than free-matrine after intravitreal injection.MAT-PLA-MS is an excellent drug delivery system.

Background The optic quality of Toric intraocular lens (IOL)-implanted eye is affected by the residual astigmatism and individual difference of corneal spherical aberration and different magnification from steep and flat axis refraction.Whether correcting Toric IOL spherical aberration can effectively improve the image quality of individual patient is a question to be studied.Objective This study attempted to collect eye parameters of cataract patients to reconstruct the customized vision model by using Zemax optical software,and to evaluate the image performance with different Toric IOL spherical aberration.Methods A prospective study was performed.Forty-five eyes of 45 cataract patients were included in Second Hospital of Hebei Medical University from August 2012 to October 2013.Several relevant parameters were measured by Pentacam,including anterior and posterior surface height of cornea,corneal thickness,curvature radius of flat and steep meridians of anterior surface astigmatism,refractive diopter and curvature radius of posterior surface.The astigmatism of anterior and posterior corneal surface was described by Matlab 4.5 software.Corneal astigmatism model were set as aspheric state,and the effective position of Toric IOL was calculated using Holladay Ⅰ formula.Customized individual model eyes were constructed by Zemax software.The contrast sensitivity function (CSF) of different spherical Toric IOLs at different spatial frequencies were calculated and compared between 300 Td light environment with 3 mm pupil diameter (photopia light) and 0.3-1.0 Td light environment with 5 mm pupil diameter (mesopia light).This study was approved by Second Hospital of Hebei Medical University ethics committee,all the patients signed the informed consent.Results The mean astigmatism power was (1.51 ± 0.36) D and (1.49 ± 0.37) D,and the mean astigmatism meridian was (101.5 ± 59.8) ° and (101.9±58.5) ° in the model eyes and cataract eyes,respectively,without significant differences between them (t=0.886,0.652;both at P＞0.05).Bland-Altman test showed a good agreement in astigmatism power and astigmatism meridian between model eyes and cataract eyes.The LogCSF values at 1.5,3.0,6.0,12.0 and 18.0 c/d spatial frequencies were significantly higher in the aspherical Toric IOL model eyes than those in the spherical Toric IOL model eyes,and the LogCSF values at various spatial frequencies were significantly higher in the Toric IOLs with spherical aberrations of-0.13 μm and-0.26 μm than those in the zero spherical aberrations in both photopia light and mesopia light (all at P＜0.05).Conclusions A precise corneal astigmatism model based on cornea high data of cataract eyes was successfully constructed through special formulas with Zemax software.Aspherical Toric IOL can compensate for spherical aberration of cornea and enhance the optic quality in individual model eye.

Background Proliferative vitreoretinopathy (PVR) is a common cause of vision loss clinically,and retinal pigment epithelium (RPE) cells play a major part in this disease.Studying the effect of traditional Chinese medicine on RPE cells are of great importance to reveal the pathogenesis and prevention of PVR,which were rarely reported.Objective This study was to study and compare the inhibition effect among curcumin,salvia miltiorrhiza and matrine on IL-1β-induced proliferation of rabbit RPE cells.Methods RPE cells at passages 3-4 were enrolled for the research and identified by transmission electron microscope.The proliferation effect of IL-1 β (2.5,5.0,10.0,20.0 μg/L) and inhibitory effect of curcumin (5,10,20 μg/ml),salvia miltiorrhiza (5,10,20 μg/ml)or matrine (100,200,400 μg/ml) on RPE cells 24,48 and 72 hours after cultivation were studied by MTT assay.The 50％ inhibitory dose (IC50) of the three medicines were analyzed by regression analysis.The use and feeding of the experimental animals were followed by the ARVO Statement.Results RPE cells isolated from the rabbit eye were in round shape and abundant in melanin;The melanin significantly decreased in the fourth generations of RPE cells.Immunohistochemistry showed that the RPE cells was positive for keratin (AE1/AE3).The proliferation rates of RPE cells were statistically different among different concentrations of IL-1β 24,48 and 72 hours after cultivation (Ftime =30.33,P =0.00;Fconcentration =9.37,P =0.00);The proliferation rates of RPE were significantly different among different time points or different concentrations of IL-1β (all at P ＜ 0.05).And the proliferation rate run up to maximum at 10 μg/L after 72 hours of cultivation.The inhibitory rates of the three medicines were statistically different among different time points or different concentrations (curcumin:Ftime =128.75,P =0.00;Fconcentration =334.05,P=0.00.salvia miltiorrhiza:Ftime =39.32,P=0.00;Fconcentration =165.57,P=0.00.matrine:Ftime =267.76,P =0.00;Fconcentration =912.34,P =0.00).The three medicines dose-dependently and time-dependently inhibit IL-1β-induced proliferation of RPE cells,with significant differences between the adjacent time points and concentrations (all at P＜0.05).The IC50 were 26.77,19.01 and 9.45 μg/ml for curcumin;33.72,23.47 and 12.56 μg/ml for salvia miltiorrhiza,570.96,352.25 and 97.50μg/ml for matrine 24,48 and 72 hours after cultivation.Conclusions The proliferation of RPE cells can be stimulated by IL-1β,and the maximal proliferation occurred with a concentration of 10.0 μg/L IL-1β.Curcumin,salvia miltiorrhiza and matrine dose-dependently and time-dependently inhibit proliferation of RPE cells induced by IL-1β.Curcumin is the best medicine to inhibit the proliferation of RPE cells.

Background Interleukin-1β (IL-1β) is an important inflammation-related factor in the initial stage of proliferative vitreoretinopathy (PVR).The previous research showed that curcumin can inhibit IL-1 β-induced proliferation of rabbit retinal pigment epithelium (RPE) cells,but the anti-inflammatory mechanism and effect of curcumin are still undefined.Objective This study was to observe the migration of IL-1β-induced rabbit RPE cells,and evaluate the function and mechanism of inhibition of curcumin on IL-1β-induced inflammation of RPE cells.Methods Cultured rabbit RPE cells of generation 4 were used in this experiment.The cells were cultured in serum-free DMEM and 0,0.1,1.0 and 10.0 μg/L IL-1β were separately added in the medium for 24 hours.The expressions of cyclooxygenase-2 (COX-2) protein and mRNA in the cells were detected by Western blot and reverse transcription PCR to determine the optimal concentration of IL-1β.The cells were divided into IL-1β group and curcumin+IL-1β group,and 1.0 μg/L IL-1 or 1.0 μμg/L IL-1 β combined with 10 μg/ml curcumin was respectively added into the medium for 24,48 and 72 hours.The cells cultured by only serum-free medium served as the control group.Hematoxylin and eosin staining was conducted for the cells to count the number of cells migrating into the injured area under the optical microscope.The relative expression levels of COX-2 protein and mRNA in the cells were detected by Western blot and reverse transcription PCR,and the relative expression levels of nuclear factor (NF)-κBp65 and inhibitor of NF-κB-α (IκB-α) protein were also detected by Western blot assay.The expression intensity and location of NF-κBp65,IκB-α and COX-2 in the cells were detected by immunochemistry.Results RPE cells just isolated from the rabbit eyes were in round shape and abundant in melanin.The melanin significantly decreased in the fourth generations of RPE cells.The shape of cells became long and narrow,and net shaped distribution.Immunochemistry demonstrated the strong positive response of RPE cells for keratin (AE1/AE3).There were (31.93 ±1.21),(36.27±2.50) and (38.33±2.40) migratory cells in the control group after 24,48 and 72 hours respectively.The number of migratory cells increased to 45.73 ± 2.30,71.13 ± 1.92 and 80.60 ± 1.71 in the IL-13 group,but obviously decreased to 13.13 ± 2.20,14.93 ± 1.10 and 12.60 ± 1.51 in the curcumin + IL-1β group.A Significant increase in the migrating cell number was found in the IL-1 β group compared with the control group and the curcumin+IL-1β group in various time points (all at P＜0.05).The relative expression levels of COX-2 protein and mRNA peaked in the 1.0 μg/L IL-1β group,so 1.0 μg/L of IL-1β was determined as the optimal concentration in the experiment.In 24,48 and 72 hours after culture,the expression levels of COX-2 protein and mRNA in the cells were significantly lower in the curcumin + IL-1β group than those in the control group (all at P＜0.05).The relative expression level reached peak in NF-κBp65 protein and lowed bottom in IκB-α proteins at 48 hours after cultured in the IL-1β group,and the reverse trend was seen in the curcumin+IL-1β group,with the significant differences between the two groups (both at P＜0.05).Immunochemistry showed that NF-κBp65 was expressed strongly in the cell nuclei and cytoplasm in the IL-1 β group and presented the weaker expression in the control group and the curcumin+IL-1 β group.Compared with the control group,the expression was weaker in IκB-α and stronger in COX-2 in the IL-1β group.In addition,the expression of IκB-α was enhanced and that of COX-2 was attenuated in the curcumin+IL-1β group in comparison with the IL-1β group.Conclusions Curcumin inhibits the movement of rabbit RPE cells induced by IL-1β.IL-1β up-regulates the expression of COX-2 by activating NF-κB signal pathway,and curcumin plays an anti-inflammatory role by blocking this pathway.

Background Choroidal neovascularization (CNV) is a common pathological basis of many ocular fundus diseases.Some treating methods are proved to be effective on CNV but there exist their own shortages.Celecoxib can inhibit experimental neovescularization.Sustained release drug of celecoxib and application approach can offer a basis for the therapy of CNV.Objective This study was to evaluate the sustained release ability of celecoxib-poly lactide-co-glycolide microsphere (CEL-PLGA-MS) in vitro and its inhibitory ability on experimental CNV in vivo.Methods CEL-PLGA-MS was prepared by Hebei Medical University and examined under the scanning electron microscope.The size of CEL-PLGA-MS was measured by Laser Particle Size Analyzer.The drugloading in vitro releasing was monitored by high performance liquid chromatograph (HPLC).Experimental CNV was induced by laser photocoagulation of retina in the right eyes of 72 male brown Norway (BN) rats and then were randomized into the CEL-PLGA-MS group,celecoxib group,blank PLGA group and PBS group.CEL-PLGA-MS with 320 μmol/L celecoxib,80 μmol/L celecoxib,blank PLGA microspheres solution and 0.01 mol/L PBS was intravitreally injected separately according to the grouping.CNV was assessed by fundus fluorescein angiography (FFA) on the 14th day after injection.The fibrovascular proliferation (FVP) thickness at photocoagulation spots was measured by OCT.The retinal pigment epithelium (RPE)-choroid-sclera sections were prepared for the histopathologieal examination of FVP.On the 7th and 28th day after intravitreal injection,the relative expression levels of VEGF mRNA and COX-2 mRNA in the photocoagulation area were detected by reverse transcription PCR (RTPCR).The use and feeding of the experimental animals were followed by the ARVO statement.Results CELPLGA-MS showed the spherical shape with the mean size of 2 467.9 nm and the drug-loading of 7.77％ and the drugrelease rate of 80.91％ in vitro for 45 days.It presented the controllable release characteristics.CEL-PLGA-MS agglomerated in vitreous body after injection.On the 14th day after intravitreal injection,the mean FVP thicknesses were (94.67±4.64),(98.56±4.72),(71.00±4.77),(50.44±3.01) μm in the blank PLGA microspheres group,PBS group,celecoxib group and CEL-PLGA-MS group,respectively,showing significant increases in mean FVP thickness in the blank PLGA microspheres group and PBS group compared with the celecoxib group and CEL-PLGAMS group (all at P＜0.01),and the CEL-PLGA-MS group appeared a lower mean FVP thickness value than the celecoxib group (P＜0.01).FFA revealed a large number of strong hyperfluorescences at the photocoagulation area in the rat eyes of the blank PLGA microspheres group and PBS group;while only weak hyperfluorescences were seen in the eelecoxib group and CEL-PLGA-MS group.Histopathological examinations verified the same results in the FVP thickness to OCT image.The relative expression levels of COX-2 mRNA and VEGF mRNA in the RPE-choroid-sclera were all significantly elevated in the blank PLGA microspheres group compared with the celecoxib group and CELPLGA-MS group both on the 7th and 28th day after intravitreal injection (all at P＜0.01).On the 7th day after injection,the relative expression levels of COX-2 mRNA were lower on the 7th day and the relative expression levels of COX-2 mRNA and VEGF mRNA were higher on the 28th day in the celecoxib group in comparison with the CEL-PLGA-MS group (all at P＜0.01).Conclusions CEL-PLGA-MSs are even in size with the spherical shape and controllable release characteristics in vitro.CEL-PLGA-MS can inhibit experimental CNV and was more durable effective than celecoxib after intravitrea] injection.