BACKGROUND: Recent studies demonstrate that erythropoietin (EPO) can protect retina from light injury, and is the mechanism related to the expression of Caspase-3 in the light-injured retinal pigment epithelium (RPE) cells. OBJECTIVE: To study the effects of EPO at different dosages on the expression of Caspase-3 in light-injured human RPE cells. DESIGN: Control observation.SETTING: Qingdao University Medical College. MATERIALS: Adult ARPE-19 cells (American Cell Culture Collection Company); DMEM/F12 mixed medium, fetal bovine serum and trypase (GIBCO Biotechnique Company); recombinant human EPO (rhEPO, Sigma Biotechnique Company); human Caspase-3 quantitative kit (Shanghai Xitang Biotechnique Co.,Ltd); Caspase-3 monoclonal antibody (American Santa Cruz Company); PV6001 immunohistochemistry kit and DAB color reagent kit (Zhongshan Biotechnology Company, Beijing).METHODS: The experiment was carried out in the Department of Pathophysiology at Qingdao University Medical College between May 2006 and January 2007. Human RPE-19 cell strain at passages 2-5 were harvested for light injury models, and the passage cells were divided into 7 groups randomly, with 4 apertures in each group:①normal control group: no light or EPO intervention;②light-injured model group: 12-hour illumination, no EPO intervention;③light-injury and EPO groups: 12-hour illumination with 10 000, 20 000 and 40 000 U/L EPO;④light-injury and 40 000 U/L EPO and AG490 group: 12-hour illumination with 40 000 U/L EPO and inhibitor of Jak2 enzyme 50 000 U/L;⑤light-injury and 40 000 U/L EPO and carbxyl-terminal modulator protein (CTMP) group: 12-hour illumination with 40 000 U/L EPO and specific inhibitor of protein kinase B enzyme 100 μmol/L.MAIN OUTCOME MEASURES: The enzyme linked immunosorbant assay (ELISA) and immunohistochemistry were used to assess the effects of rhEPO at the different doses on the expression of Caspase-3 in light-injured human RPE cells. RESULTS: Caspases-3 was not expressed in RPE cells of the normal control group and was positively expressed in the nucleus of RPE cells of the light-injured model group, showing a specific brown-yellow staining. Expression of Caspase-3 was gradually decreased in every rhEPO group with increase of EPO concentration, with the weakest expression in 40 000 U/L rhEPO group. The effects of EPO on Caspase-3 expression were strongly inhibited in light-injury+ 40 000 U/L EPO +AG490 group and the expression was positive in light-injury +40 000 U/L EPO+CTMP group, which was slightly weaker than light-injured model group. CONCLUSION: The rhEPO can reduce the expression of Caspase-3 in the light-injured human RPE cells, and one of the possible mechanisms is the inhibition of light-injured RPE cell apoptosis by the rhEPO.

Anthracosis ; complications ; Cerebrovascular Disorders ; complications ; Cohort Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies

ObjectiveTo investigate the expression of hypoxia inducible factor 1 (HIF-1α) in rats' retinae during the embryonic and earlier postnatal period.MethodsThe retinal expression patterns of HIF-1α protein and mRNA of embryonic day 12 (E12), E16, E20, and postnatal day 1(P1) and P5 rats were determined by immunohistochemical staining and reverse transcription polymerase chain reaction (RT-PCR).ResultsHIF-1α protein was detected in the neural epithelial layer and the pigment epithelial layer at all those 5 timepoints, with higher expression in the ganglion cell layer and the inner plexiform layer, and seems limited to the ganglion cell layer when retina became more mature. Embryonic rat retina had higher expression of HIF-1α protein and mRNA than postnatal retina, the difference was significant (P<0. 01). ConclusionThe expression of HIF-1α in rats' retinae differs from embryonic to earlier postnatal stages.

Adult ; Female ; Humans ; Maxillofacial Abnormalities ; Nose ; abnormalities

Objective To explore the clinical.significance of serum anti-lysosomal associated membrane protein-2 (LAMP-2) antibody in the pathogenesis of anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AASV) by investigating the relationship of its levels and AASV.Methods Sera from twenty patients with AASV and twenty healthy controls were collected.Serum anti-LAMP-2 antibody was detected using commercial enzyme linked immunosorbent assay (ELISA) kits.Anti-LAMP-2 antibody levels between the two groups were assessed using the t test,the correlation between anti-LAMP-2 antibody levels and erythrocyte sedimentation rate (ESR),C-reactive protein (CRP) was assessed by Spearman's rank correlation test,the correlation between anti-LAMP-2 antibody levels and the Birmingham vasculitis activity score(BVAS),hemoglobin (Hb),albumin (Alb) was assessed by Pearson's rank correlation test.Results ① The serum level of anti-LAMP-2 antibody in patients with AASV [(3714±1446) pg/ml] was higher than that in the healthy controls [(174±43) pg/ml] (t=10.94,P＜0.05).The serum level of Hb [(99±30) g/L] and Alb [(27±5) g/L]in patients with AASV was lower than that in healthy controls [(138±14) g/L,(44±3) g/L] (t=5.27,t=13.04,P＞0.05).② The level of anti-LAMP-2 antibody in AASV was positively correlated with BVAS (r=0.669 9,P＜0.05),and was not found elevated compared with ESR,CRP,Hb and Alb (P＞0.05).Conclusion ① AntiLAMP-2 antibody is involved in the pathogenesis of AASV.② Anti-LAMP-2 antibody is correlated with the activity of AASV,it may be an indicator of AASV disease activity.

Objective To establish a specific HPLC method and a standard fingerprint for quality control ofZiziphi Spinosae Folium.Methods The samples were separated on a Kinetex C18 column (100 mm × 2.1 mm,2.6 μm) by gradient elution at the flow rate of 300 μL/min using acetonitrile and 0.1％ aqueous formic acid as the mobile phase.The column temperature was 30℃.The detection wavelength was 225 nm and the sample size was 10 μL.The fingerprint evaluation software (2012 edition) for Chinese materia medica (CMM) was used to evaluate the similarity of the 12 batches of samples.Results There were 13 characteristic peaks identified in the characteristic spectra ofZiziphi Spinosae Folium samples.Peak 6 was Rutin.The similarities of 13 batches ofZiziphi Spinosae Folium samples were proved to behigher than 0.850.Conclusion The method is available with a good reproducibility and accuraty which can control the quality standards effectively.

Yttrium vanadate:europium nanoprobes (YVO4∶Eu NPs) with good fluorescence properties and water solubility were synthesized by solvent thermal method.Due to the overlapping of the excitation spectrum of YVO4∶Eu NPs and the absorption spectrum of tryptophan, fluorescent internal filter effect (IFE) occurred, in which YVO4∶Eu NPs were the fluorophore and tryptophan was the absorber, leading the fluorescence of YVO4∶Eu NPs was quenched.Therefore, a new method for the determination of tryptophan was established by using fluorescent YVO4∶Eu as nanoprobes based on IFE.Some experimental parameters, such as the adding amount of YVO4∶Eu NPs, pH value of the reacting solution, and reacting time, were investigated.Under the optimum reaction conditions, the linear range of the method was 4.0×10-6-4.0×104 mol/L and the detection limit was 1.0×10-6 mol/L (3σ).The content of tryptophan in the soy sauce was determined with the recovery of 95.2% and 97.3%.This method is simple, rapid, sensitive and accurate.

Aim This study aimed to assess the protection of recombinant human erythropoietin (rhEPO) in light-induced injuries in human retinal pigment epithelial(RPE)cells by researching the inhibition of rhEPO for apoptosis in human RPE cells by light-induced injuries.Methods Cultured human RPE cells were exposed to light of 8 w (2 000?500) lux for 12hours,then the culture were stopped at 24 hours after 12hours light stimulation. The effect of inhibiting apoptosis of rhEPO was detected by AnnexinV-flunorescein isothiocyanate/Propidium iodium labeling and flow cytometry. The enzyme linked immunosorbant assay(ELISA)and immunocytochemical staining were used to assess the expressions of caspase-3 and Bcl-2 treated by different doses of rhEPO in light-induced injury on human RPE cells and research the protective mechanism of rhEPO by adding AG490(the special inhibitor of Jak2).Results There was a obviously increased effects on inhibiting apoptosis in every rhEPO group, which was the most conspicuous in 40 IU?ml-1 rhEPO group,and the value was (4.93?1.45)?ml-1. The decrease of expression of caspase-3 was most obvious in 40 IU?ml-1 rhEPO group, and the value was (0.125?0.029) ?g?L-1. The increase of expression of Bcl-2 was the most obvious in 40 IU?ml-1 rhEPO group and the value was 168.21?3.87. But these effects on inhibiting apoptosis in rhEPO group were restrained by adding AG490, the value of apoptosis was (11.29?2.11)?ml-1 and the density of caspase-3 increased to (0.362?0.042) ?g?L-1,the expression of Bcl-2 dropped.Conclusion It is suggested that rhEPO can inhibit the apoptosis of human RPE cells in the light-induced injuries and inhibit the expression of caspase-3 and up-regulate the expression of Bcl-2, so rhEPO can protect the light-induced injuries for human RPE cells. Its protective mechanism is accomplished principally by the pathway of combining EPO with EPOR ,then the combination activates Jak2.

Objective To investigate the growth and expression of PDGF of VSMCs in response to stimulation of autologous serum and mechanical injury. Methods An vitro model simulating the condition as possible as that after PTA.PDGF of every medium sample from every group was detected by ELISA,and the values of MTT of every cellular sample was measured by MTT to show the growth and proliferation of every group. Results After stimulation by autologous serum and mechanical injury,SMCs of the experimental group showed the value of MTT increasing,but SMCs in control group reached on 3rd day.At the same time,the expression of PDGF also increased gradually,obtaining peak gradually up to peak on day 4/5 nearly 2.0-fold as much as that of SMCs in the control group. Conclusions After on the 5th day,stimulation with autologous serum and mechanical injury,VSMCs of rabbit showed the stronger ability of growth/proliferation,and autocrine of PDGF also increased gradually,reaching peak on 4-5 d,probobly simulating to those in vivo.

Objective:To investigate the efficacy and the clinical value of magnetic resonance elastography(MRE) in diagnosis of hepatic fibrosis with Meta-analysis, and to provide basis for clinical treatment of hepatic fibrosis.Methods:The studies published before February 2, 2017 about MRE and staging of hepatic fibrosis in Chinese or English were retrived in the databases including PubMed, EMBase, Web of Science, Cochrane Library,CNKI, CBMDisc,VIP, Wanfang data, and supplemented by manual retrieval for relevant literatures.The inclusion and exclusion criterions were used to select and extract the literatures.The literatures qualitie were valuated based on QUADAS-2 tool.The sensitivity(SEN), specificity (SPE), diagnostic odds ratio (DOR), positive likelihood ratio (+LR), negative likelihood ratio (-LR) on the groups of F0 vs F1-F4,F0-F1 vs F2-F4,F0-F2 vs F3-F4, F0-F3 vs F4 and heterogeneity were combined and tested with Stata software respectively.HSROC and AUROC were also implemented.Results:A total of 1 332 studies were searched, and 22 were included.21 of them were in English and 1 in Chinese.The results of Meta analysis showed that the SENp, SPEp, +LRp,-LRp, DOR and AUROC in F0 vs F1-F4 group were 88.8%(85.0-91.7),95.9%(91.5-98.0),21.435(10.215-44.979),0.117(0.086-0.159),183.187(72.533-462.650) and 0.96(0.94-0.98) ,respectively;the SENp, SPEp, +LRp,-LRp, DOR and AUROC in F0-F1 vs F2-F4 group were 93.3%(89.2%-35.9%), 94.1%(90.2%-96.5%),15.839(9.344-26.848),0.072(0.044-0.117),221.224(100.980-484.648) and 0.98(0.96-0.99),respectively;the SENp, SPEp, +LRp, -LRp, DOR and AUROC in F0-F2 vs F3-F4 group were 92.9%(88.9%-95.5%),94.6%(91.2%-96.8%),17.348(10.496-28.671),0.075(0.048-0.119),230.434(111.482-476.317)0.98(0.96-0.99), respectively;the SENp, SPEp, +LRp,-LRp, DOR and AUROC in F0-F3 vs F4 group were 97.7%(93.0%-99.3%),93.2%(90.3%-95.2%),14.337(9.910-20.742),0.025(0.008-0.075),580.405(144.871-2325.307) and 0.98(0.96-0.99),respectively.Conclusion:MRE,as a new and noninvasive imaging method, has high diagnostic value in all stages of hepatic fibrosis, which can provide a reliable reference for clinical precise treatment of hepatic fibrosis.