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.

BACKGROUND: Hypoxia inducible factor-1 α(HIF-1 α) is not only related to physiological reaction of hypoxia,but also takes part in normal embryonic development.OBJECTIVE: To study the alteration of HIF- 1 α in retina during rat development.DESIGN,TIME AND SETTING: Randomized contrast animal study,which was performed in the Shandong Provincial Molecular Virus Key Laboratory,Medical College of Qingdao University between January and September 2007.MATERIALS: Adult Wistar rats,nulliparity,clean grade,and weighing 200 250 g were used in this study.METHODS: Male and female rats were caged as the ratio of 1 : 1.Embryos were obtained at 12-day,16-day,and 20-day pregnancy.Eyeballs were obtained from newborn rats by anesthesia at 1-day,5-day,10-day,and 12-month birth.Retina was separated and made into paraffin section.MAIN OUTCOME MEASURES: Expressions of HIF 1 α protein and HIF-1 α mRNA in retina were measured by immunohistochemistry and semiquantitative reverse transcription-polymerase chain reaction at various dine points of embryonic development.RESULTS: HIF-1 α positively expressed at stratum neuroepitheliale retinae and purpurogenous membrane in the embryonic phase.Additionally,HIF-1 α still positively expressed at stratum neuroepitheliale retinae and purpurogenous membrane,especially in ganglionic cells and inner plexiform layer,in early development.With the gradual development,the positive expression was mainly located at stratum ganglionare retinae.HIF-1 α protein and mRNA expressions were the highest in the embryonic phase,lower in the development,and the lowest in the adult period.There were significantly differences among these three phases (P ＜ 0.01).CONCLUSION: HIF-1 α decreases gradually in retina and its expression is mainly located at stratum ganglionare retinae.

Virginiamycin acetyltransferase D (VatD) plays a vital rule in streptogramins resistance by chemically inactivating streptogramin A. Therefore, it is desirable to discover novel small molecular weight inhibitors of VatD via state-of-the-art virtual screening techniques. This "cocktail" strategy by combining VatD inhibitor with streptogramins may provide new therapeutic opportunity for resistant bacteria infections. Structure-based virtual screening method (molecular docking) was applied to rank and score a chemical database containing 300 000 commercially available compounds against the VatD substrate binding site. Twenty six out of the 200 top scored compounds from the docking calculation were selected and submitted to the VatD enzymatic inhibition assay. The plasmid pRSET B/vatD was constructed and transformed into E.coli (trxB) host cells for over-expression, and VatD enzyme was purified and validated by showing acetyltransferase activity to Virginiamycin M1. Three out of these 26 tested compounds showed enzymatic inhibition on VatD with IC50 168.6, 91.0 and 55.2 μmol·L-1, separately. Other compounds could not be dissolved in the system and/or had little effect on the enzyme (IC50>200 μmol·L-1). To our knowledge, it is first time that small molecular weight organic compounds were identified as VatD inhibitors. It is expected that the VatD inhibitors identified at present study could serve as lead compounds for the further development of the novel therapeutic agents to overcome streptogramins resistance.

BACKGROUND: Basic fibroblast growth factor (bFGF), a kind of polypeptide growth factor possessing multifunctional biological activities,can protect neurons and promote the growth of nerves. It has been corfirmed that bFGF has therapeutic effects on retina ischemia/reperfusion injury (RIRI).OBJECTIVE: To establish RIRI model and analyze the effects of bFGF on cellular apoptosis of retina and the expression of regulatory gene protein.DESIGN: Randomized grouping and validating trial.SETTING: Department of Ophthalmology, the Affiliated Hospital of Medical College of Qingdao University.MATERIALS: The experiment was conducted at the Research Laboratory of Pathology, Department of Ophthalmology, Medical College of Qingdao University, from April 2002 to December 2003. Twenty-eight healthy Wistar rats were enrolled in this experiment. Four rats were randomly chosen for normal control group, the left eyes of the other 24 rats were set as normal saline control group, and the right eyes were set as bFGF group.METHODS: Normal saline control group and bFGF group adopted the rat RIRI models established by transiently elevating intraocular pressure. Normal saline of 12 μL was injected into the vitreous cavity of the left eyes of the rats in normal control group. 12 μL bFGF was injected into the vitreous cavity of the right eyes of the rats in bFGF group, 4 rats once. No administration was given in normal control group. The expression of apoptotic cells was detected and apoptosis indexes were calculated with the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) method and immunohistochemical staining method at the 1st, 6th,12th, 24th,48th and 72nd hours after reperfusion and ischemia for 1 hour.MAIN OUTCOME MEASURES: ① The detection results of apoptotic cells in situ of retina tissuesat different time points after reperfusion. ②The expression of Fas and caspases-2 in retina tissues at different time points after reperfusion.RESULTS ① Comparison of apoptosis indexes of retina tissues at different time points after ischemia reperfusion: There were no apoptotic cells in the retina tissues of the rats in normal control group. As compared with those in normal saline control group, apoptosis indexes in bFGF group were significantly decreased at ischemia 1 hour and reperfusion 1, 6, 12, 24, 48and 72 hours, especially at the 12th, 24th and 48th hours after reperfusion (t =5.362-5.595, P ＜ 0.05). ② The change of Fas expression at different time points after ischemia reperfusion: There was hardly any Fas expression in normal control group. As compared with that in normal saline control group, Fas expression in bFGF group was significantlydecreased at ischemia 1 hour and reperfusion 1, 6, 12, 24, 48 and 72 hours, especially at the 6th, 12th and 24th hours after reperfusion (t=3.954-9.327, P ＜ 0.05). ③The changes of caspase-2 expression at different time points after ischemia reperfusion: There was no caspase-2 expression in normal control group.Compared with that in normal saline control group, the number of caspase2 positive cells in bFGF group was significantly decreased at the 6th,12th,24th, 48th and 72nd hours after ischemia for 1 hour and reperfusion (t=4.125-15.641, P ＜ 0.05).CONCLUSION: bFGF can significantly inhibit the expression of apoptosis gene Fas and caspase-2 in the ischemia and reperfusion of retina, thus reducing cellular apoptosis of ganglion cells and exerting therapeutic effects on the ischemia and reperfusion of retina.

Aim To explore the effects of tanshinone IIA on cell proliferation via G protein-coupled estrogen receptor inductive and regulative pathway in typical es-trogen receptor and G protein-coupled estrogen receptor positive T47D breast cancer cells. Methods The pro-liferation rate of T47 D cells influenced by tanshinone IIA was analyzed by MTT assay. G protein-coupled es-trogen receptor agonist G1 and GPER antagonist G15 were employed as tools. GPER SiRNA was applied to build GPER gene silence T47D cells. GPER expres-sion influenced by tanshinone IIA was measured by Western blot. Results The proliferation rates of T47D cells treated with 1 × 10 -5 mol·L-1 - 1 × 10 -7 mol· L-1 of tanshinone IIA were decreased significantly. Such effects could be attenuated by G1 or enhanced by G15 . Growth of GPER SiRNA transfected T47 D cells were significantly inhibited by 1 × 10 -5 mol·L-1 - 1 × 10 -7 mol·L-1 of tanshinone IIA treating. Result of Western blot showed that tanshinone IIA at 1 × 10 -5 mol· L-1 and 1 × 10 -6 mol · L-1 could induce de-crease of GPER protein expression in T47D cells. Conclusions Tanshinone IIA shows inhibitory effects on proliferation rate of T47 D breast cancer cells via GPER pathway. Tanshinone IIA could perform regula-tive function on GPER expression level in target cells.

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 put forward a design of the software of brainwave therapeutic apparatus based on the MCU. Methods Based on principle of electroencephalogram music treatment, several sound synthesis techniques were analyzed and contrasted. A simple and easily-realized one was selected and analyzed in detail. Finally, the output of brainwave therapeutic apparatus was tested and errors were analyzed. Results The selected scheme can effectively generate brainwave music. Conclusion According to the scheme, a low-cost and reliable brainwave therapeutic apparatus can be made by using the MCU.

To popularization and improvement of on-the-spot rescueability is a vital means to reduce the accidental disability rate and the mortality rate,which is also a prospective concept of health.To popularize on-the -spot rescue knowledge and to improve self and mutual medical aid is a strongly public welfare and is the manifestation of social responsibility as well as respect for life.Based onthe investigation of on-the-spot rescue ability among three higher vocational college students in Shaanxi,this paper pointed out that the popularity rate of on-the -spot rescue knowledge was low among higher vocational college students,which no longer kept pace with the demand of social development and occupational requirement.It also analyzed the importance and urgency of improving higher vocational students'on-the-spot rescue ability from the perspective of social ethics.Strengthening the propaganda and training self-rescue skills might be a possible way to enhance students' on-the-spot rescue ability.