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.

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.

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

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.

Objective To observe the curative effects of octreotide on gastrointestinal cancer and its influenee on the serum level of IGF-1.Methods 33 patients diagnosed as advanced gastrointestinal cancer were randomized into 2 groups.15 cases in therapy group received octreotide 400μg/d subcutaneous injection or intravenous injection,other patients were taken as control.Curative effects of octreotide and serum level of IGF-1 on different time were observed before and after therapy.Results 4 cases in therapy group kept stable condition during treatment,all 18 cases in control group deteriorated gradually.The median survival time of octreotide therapy was 6.9 months,longer than that of control group(2.3 months)(P＜0.05),the feeling of well being in therapy group was reported a remarkable improvement.Serum level of IGF-1 decreased obviously after octreotide injection(P＜0.05),but no significant difference in control group.Conclusion Octreotide can prolong the survival duration and improve living quality of patients with advanced gastrointestinal tumors.The mechanism of antitumor is probably through suppression of IGF-1.

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 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.

Background Erythropoietin (EPO) was proved to be express in hematopoietic tissue and nervous system and play the effects of stimulating blood cell production and protecting nervous tissue.Researches showed that EPO is expressed in the embryon brain of animal.However,whether EPO exist in nervous-derived retina and its action on retina with the development is concerned. Objective This research was to investigate the expression of EPO during the embryonic development period of rat retina and explore the role of EPO in retina development process.Methods Clean Wistar rats with pregnancy for 12 days,16 days and 20 days were collected,and the embryonic 12-day rats (E12 d,5 rats),embryonic 16-day rats (E16 d,5 rats) and embryonic 20-day rats ( E20 d,5 rats) were obtained by caesarean operation,and 5 12-month W istar rats were used as controls.The rats were sacrificed by cervical dislocation and the retinal sections were prepared in the different-embryo-phase (12 d,16 d,20d) and growth phase.The expression of EPO protein and mRNA in rat retina was detected by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR),respectively.The feed and use of the animals followed the Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results EPO was positively expressed in the cytoplasm and nuclei in the neuroepithelial layer and pigment epithelium of every-embryo-phase rats but only in retinal ganglion cell layer in 12-month-old rats.The gray scale values of EPO expression in retina were 105.55±10.35,99.35± 8.71,83.27± 7.84and 30.30± 3.80 in E12 d rats,E16 d rats,E20 d rats and 12-month-old rats respectively with a statistically significant difference (F=76.13,P＜0.01 ).RT-PCR revealed that the relative values of EPO mRNA expression in retina were 0.876±0.10,0.861 ±0.09 and 0.256±0.03 in E16 d rats,E20 d rats and 12-month-old rats respectively,presenting a elevated value in embryonic rats compared with adult rats ( P =0.00).Gel imaging deletion showed that the A value of EPO amplification products was highest in E16 d rats and lowest in adult rats.Conclusions The expression of EPO appears a high to low fashion during the embryonic development of Wistar rats,which is closely associated with the developing procedure of retina.

Background Pigmentary glaucoma and pseudoexfoliation glaucoma are characterized by pigment dispersion in trabecular meshwork,and the dipersitional pigment probably contributes to the resistance of aqueous outflow pathway,irreversible damage of the trabecular meshwork and the remodeling abnormality of extracellular matrix.Researches determined that contents of transforming growth factor-β (TGF-β) in the aqueous humor are increased in glaucomatous eyes.However,the effects of TGF-β and fibronectin (FN) in the trabecular meshwork cells (TMCs) acted by iris pigmentary particles still are not elucidated.Objective This study was to investigate the effects of iris pigment particles on TGF-β1 and FN expression in bovine TMCs (BTMCs) cultured in vitro.Methods BTMCs were cultured in vitro by explant culture method and identified by morphological evaluation.The third generation of BTMCs were divided into normal control group and pigment group,and 100 μ1 PBS and 100 μl iris pigment suspension (final concentration of 1 × 107 particles/ml) were added into the medium for 24 hours,respectively.The expressions of TGF-β1 mRNA,FN mRNA and their proteins in the BTMCs were assayed by real-time fluorescence quantitative PCR and ELISA,respectively.Results Cultured cells grew well and showed the fusiform,polygon and dendritic like in shape,with pigmented and round nuclei.The relative expression levels of TGF-β1 mRNA and FN mRNA in the cells were 2.98±0.27 and 0.36±0.10 in the iris pigment group,which were significantly higher than 1.00±0.00 and 1.00±0.00 in the normal control group (t =12.68,10.60,both at P =0.00).The concentrations of TGF-β1 protein and FN protein in the cell suspension were (156.60±9.74)ng/L and (59.29±15.79)mg/ml in the iris pigment group,showing significant differences in comparison with (65.46 ± 14.24) ng/L and (102.10 ± 12.14)mg/mlin the normal control group (t=9.15,P=0.00;t=3.72,P=0.02).Conclusions The expression of TGF-β1 is up-regulated and that of FN is down-regulated in BTMCs cultured by iris pigment,inferring that TGF-β1 and FN participate in the pathogenesis and development of pigmentary and pseudoexfoliation glaucoma.

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.