Objective To optimize extraction technology of flavonesingredients from Herba Epimedii by uniform design. Methods Ultraviolet spectrophotometry was adopted to determine the content of total flavonoids. Content of epimedin A, epimedin B,epimedin C,icariin and baohuosideⅠ were determined by HPLC. U10?( 108 ) uniform design was used to conduct the multiple comprehensive evaluation of six ingredients and comprehensively analyze the influence of the concentration and amount of ethanol, extracting time on extraction of flavonesin gredients from Herba Epimedii. Results The uniform design experiment showed that 15-fold weight of 60% ethanol,extracting 2 times and each time with 165 min were the optimum extraction condition. Conclusion The method is easy and reasonable to handle,has stable and reliable results,and good repeatability and feasibility.It can be applied in industrial production.

Objective To investigate the satisfaction of clinicians on clinical research associates (CRA)and its influencing factors,for the purpose of providing rationalized proposals on education of CRAs.Methods 141 clinicians were randomly sampled from tertiary hospital for questionnaire survey,using the 5-point Likert scale.The survey covers 4 levels,i.e.,the work attitude,professional knowledge and ability,communication skills,and project management capabilities,as well as 14 dimensions.Data processing and statistics were analyzed using SPSS 15.0 software.The attribute characteristics of the investigation subjects were analyzed using x2 of the contingency table,along with analysis of its correlation with the general satisfaction on CRAs.Results The mean values of the 12 indicators range 2.28 to 3.75,with low satisfaction in general.Among these indicators,satisfaction of the service attitude of the CRA,and of their familiarity with the pilot program and CRF completion axe the highest,respectively,74.04％and 61.70％.Satisfaction of the rest 10 indicators falls below 50.00％.The chisquare analysis showed no association between satisfaction and gender,education,job titles.The satisfaction is different(P＜0.05)between those trained and those not,while there exists a significant differences(P＜0.01)between those participating in different number of tests.Conclusion Clinicians have a low satisfaction on CRAs.It is recommended to strengthen the training,establish a CRA occupation certification system,and to strengthen the clinicians' emphasis and competency of clinical trials.These actions will normalize the industry of clinical trials and improve the level of clinical trials in China.

ObjectiveTo determine the source of IL-6 in detached retina and the change of IL-6 level in detached and reattached retina.MethodsRetinas of SD rat were examined after subretinal injection of 1.4% Healon GV at different period of time, and the level of IL-6 in detached and reattached retina were detected by radio-immune histochemistry method. Wax-embeded sections were labeled with IL-6 antibody to determine the location of IL-6.ResultsDetached retina with normal vitreous and inner limiting membrane could only induce the subretinal fibrosis. This kind of fibrosis reached to its peak at 10th day and then remolded with time. Retinal pigment epithelial (RPE) cell, Muller cell, endothelial cell, glial cell were labled with IL-6, and the level of IL-6 in neuro-retina reached to its peak at 3rd-4th day and then downed to normal within a few days. The level of IL-6 in reattached retina was lower than in detached retina. The expression of IL-6 in RPE of detached area was stronger than in attached area.ConclusionIL-6 takes active part in wound healing process induced by the separation of RPE and neuro-retina. Reattachment can lower the expression of IL-6 in retina.

ObjectiveTo investigate the expression of integrin β1 during retinal detachment and reattachment of rabbits.Methods24 rabbits were used to make retinal detachment and reattachment model by using hyaluronidase and micropipette. The expression of integrin β1 were observed with hybridization in situ.ResultsThe expression of integrin β1 in reattached retina was lower than that in detached retina.ConclusionRetinal reattachment may inhibit the development of proliferative vireoretinopathy.

ObjectiveTo investigate the possible effect of interleukin-1β(IL-1β)in experimental retinal detachment,and the role in proliferation.MethodsThe experimental retinal detachment and reattachment in different time were made using Sprague-Dawley rats. The expression of proliferating cell nuclear antigen (PCNA) in neuro-retina was tested with flow cytometry at different time. IL-1β in neuro-retina were analyzed by labeling with polyclonal IL-1β antibody. The level of IL-1β in neuro-retina were tested with radio-immune method. IL-1β antibody 1000 ng,IL-1Ra 20 ng were injected in the subretinal space of some rats before PCNA reached to its high point respectively,and the expression of PCNA of them were compared with that of control which were injected with 0.01 M PBS.ResultsIL-1β expressed in Muller cell,astrocyte,vascular endothelial cell in neuro-retina and reached to its peak at the 7th day after detachment,then declined and continued at a low level as long as the retina detached.The expression of PCNA began at the second day after detachment, and reached a maximum at about 10 days after,then declined and continued at a low level as long as the retina detached. IL-1βantibody and IL-1Ra could restrain the expression of PCNA.ConclusionProinflammatory cytokine IL-1β is the key factor in proliferation of experimental retinal detachment.

Objective To investigate the protective effects of imipramine on alveolar epithelial barrier functiou in mice against LPS-induced acute lung injury (ALI),and explore the possible mechanisms.Methods Total of 32 SPF male Balb/c mice were randomly (random number) divided into four groups:control group,Imipramine group,LPS group,LPS + Imipramine group.To establish an animal model of ALI,mice were administered intraperitoneally with LPS in 20 mg/kg.Mice were treated with imipramine in 25 mg/kg 30 min prior to LPS administration.FITC-FD4 was administered in mice via the tail vein with FITC-FD4 10 min before mice sacrificed under anesthesia at 12 hours after LPS administration,then bronchoalveolar lavage fluid (BALF) and lung tissue were obtained.HE staining was used to observe histopathological changes,and pathology scores;lung tissue wet-to-dry weight ratio and BALF/serum FD4 ratio were used to assess pulmonary edema and alveolar epithelial permeability.Real-time PCR,western blot and immunochemistry were employed to detect the mRNA expressions and protein levels of Occludin,Claudin-4 and ZO-1.Data were analyzed with SPSS 16.0 software,one way analysis of variance (ANOVA) was used to compare multiple sets of variables,the intergroup comparisons were analyzed by the least-significant-difference (LSD) tests with P ＜ 0.05 for the statistically significant difference.Results Compared with LPS group,LPS + lmipramine group had a statistically significant decrease in pathological score [(9.22 ± 0.21) vs.(11.23 ± O.55),P ＜ O.05);the wet-to-dry weight ratio in LPS + Imipramine group was less than that in LPS group and the difference was statistically significant (P ＜ 0.05);compared with LPS group,the ratio of BALF/serum FD4 in LPS + Imipramine was less and the difference was statistically significant (P ＜ 0.05);compared with LPS group,the mRNA expressions and protein levels of Occludin,Claudin-4 and ZO-1 in LPS + Imipramine group were significantly increased (mRNA:Occludin:P ＜ 0.05;Claudin-4:P ＜ 0.05;ZO-1:P ＜ 0.05 . western blot:Occludin:P ＜ 0.05;Claudin-4:P ＜ 0.05;ZO-1:P ＜ 0.05).Immunochemistry showed that Occludin and Claudin-4 were present mainly in alveolar epithelial cell membrane,Z0-1 was found mainly in cytoplasm of alveolar epithelial cell.In control group and Imipramine group,tight junction proteins were obviously expressed.Compared with control group,protein levels in LPS group were significantly decreased (Occludin:P ＜ 0.05;Claudin-4:P ＜ 0.05;ZO-1:t =6.59,P ＜ 0.05);compared with LPS group,the tight junction proteins in LPS + Imipramine group were significantly increased (Occludin:P ＜ 0.05;Claudin-4:P ＜ 0.05;ZO-1:P ＜ 0.05).Conclusion The protective effects of imipramine on alveolar epithelial barrier function by up-regulating tight junction proteins expression in murine LPS-induced ALI.

Background Subretinal transplantation of retinal pigment epithelium (RPE) cells for the treatment of age-related macular degeneration (AMD) have accelerated the drive to develop xeno-free cultivation system that support the rapid differentiation of human embryonic stem cells (hESCs) into ES-RPE cells.Objective This study was to report a modified xeno-free culture system and method for accelerating derivation of hESCs to differentiate into RPE cells.Methods This study was approved by Ethic Committee of Zhongshan Ophthalmic Center.HESC H1 line was cloned and cuhured in Vitronectin XFTM-coated 6-well dish with xenogenetic-free medium.Cells were cultured in 50 ng/ml noggin,10 ng/ml DKK-1 and 10 ng/ml insulin like growth factor-1 (IGF-1) medium for 2 days,and then the concentration of noggin was decreased to 10 ng/ml and 5 ng/ml basic fibroblast growth factor (bFGF) and cultured for the following 2 days.Sequentially,noggin and bFGF were removed and cultured for 2 days.Finally,1 μmol/L CHIR99021 was added in medium for 6 days.Morphological changes in the progress of ESCs differentiation into RPE were observed by Living Cell Imaging System.The expression of Mitf and RPE65,RPE cellsspecific markers,in the cells were detected by immunofluorescence technique,and the relative expression levels of RPE cells-specific marker mRNA were assayed using real time fluorescent quantitation PCR.Results Polygonalshape monolayer cells which contained pigments were initially observed at day 14 after cultured with the cobblestonelike arrangement.Mitf and RPE65 were strongly expressed in the hES-derived RPE cells 35 days after induced,showing red fluorescence,and the cells presented hexagonal shape at cultured day 60 with numerous pigment granules in cytoplasm.Compared with before differentiation,the expression levels of Mitf mRNA in hES-RPE cells increased by (3.43±2.77) folds and (8.91 ± 2.83) folds,and the expression levels of RPE65 mRNA increased by (14.60 ± 3.94) folds and (87.16 ±9.32) folds at day 7 and day 14 after differentiation,respectively (all at P＜0.05).Conclusions A defined xeno-free culture system is successfully established by adding niacinamide,DKK-l,noggin,IGF-1 and CHIR99021 in xeno-free medium,and this system can accelerate the derivation and differentiation of hESCs into RPE-like cells.

Chitosan-carboxymethyl-chitosan complex film was prepared by freeze drying. Some tests in vivo and in animal were employed, in order to evaluate it on biology. All results indicated that the film has not only good surface compatibility but also good structural compatibility. It can be more suitable for GTR technology.
Animals ; Biocompatible Materials ; chemical synthesis ; pharmacology ; Chitin ; analogs & derivatives ; Chitosan ; Materials Testing ; Membranes, Artificial ; Rabbits ; Rats ; Skin Irritancy Tests

Objective To assess left ventricular(LV)twisting parameters in different layers with velocity vector imaging(VVI)in both healthy subjects and premature ventricular contraction(PVC)bigeminy from right ventricle(RV)patients,and investigate the rules of LV myocardium dynamics in sinus rhythm and PVC from RV.Methods Ten patients with PVC bigeminy from RV and twelve healthy subj ects underwent the examination of two-dimensional ultrasound,in which the standard LV mitral valve(MV)and apical short-axis views were acquired.The twisting angle(TA)and untwisting starting time(UST)were measured.Results In healthy subjects myocardium of LV MV short-axis twisted in clockwise,while the myocardium of LV apical short-axis twisted in counter-clockwise.TA of endocardium was higher than that of epieardium(P＜0.01).In PVC bigeminy from RA patients,their TA during sinus contraction was lower than that of healthy subjects both in MV and apical short axis respectively(P＜0.05),and UST was earlier(P＜0.05).In PVC the TA curve is bidirectional.and in early systolic TA twisted reversely compared with that of healthy subjects and sinus contraction in PVC patients.Conclusions Twisting parameters could be used to assess myocardium dynamics during different ventricular mechanical activation.VVl was a novel tool to analyze myocardium twisting.

Objective To evaluate the immediate impact of radiofrequencycatheter ablation on left atrial (LA)volumes and function by velocity vector imaging(ⅤⅥ)and compare the LA function with and without atrial fibrillation after the operation.Methods Ten consecutive patients with paroxysmal AF were studied at baseline and within 3 days after ablation.Ten consecutive patients with persistent AF were studied within 3 days after ablation,in sinus rhythm.Ten patients with normal ventricular function were included in the study.ⅤⅥ was performed to assess LA sizes and strain,strain rate,velocity of the septum,lateral wall and the atrial roof from the apical four-chamber view.Results In patients with paroxysmal AF,global and regioal LA function was not significantly impaired after the ablation procedure.Subgroup analysis demonstrated that there were no significant difference in LA function betwwen patients with paroxysmal AF and control subjects,but the indexed LAVmax was significantly larger in all AF patients compared with control subjects.The global function of LA,including LAEF,LAaEF and LA expansion index significantly decreased in persistent AF patients.By contrast,the LA septal strain,velocity and lateral strain were lower than controls.Conclusions Radiofrequency catheter ablation has no influence on LA function assessed by ⅤⅥ within three days after the operation for patients with paroxysmaI AF.The function of persistent AF patients is absolutely lower than control subjects.