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.

0.05). Conclusion Air pollution might have effect on pupils' non-special immunity.

Objective　To investigate the potentiality of adhesion of S. pn to type Ⅱ pneumocyte (A549) in inducing tyrosine phosphorylation of A549 cell proteins and the function subcomponents involved in. Methods　Labelled S. pn with FITC for the observation of kinetic characters of adhesion in vitro; The tyrosine phosphorylation of A549 cell proteins induced by S. pn was examined by immunohistochemistry and ELISA. Results　The adhesion of S. pn to A549 cells was in a time-and dose-dependent fashion suggesting the specific interaction; Tyrosine phosphorylation induced by adhesion was dose-and time-dependent and was mediated by the surface proteins of S. pn. Conclusion　S. pn adheres to A549 cells and triggers the signal cascade in which the surface proteins of S. pn play a vital role.

Objective To observe the preemptive analgesia effects of katamine and elonidine, and to find out the influence of preemptive on stress responses. Methods 36 patients with hysteromyoma undergone hysteromyomec-tomy were randomly assigned to three groups (n = 12 each group) :group Ⅰ , control group, without preemptive analge-sia,the patients in control group were given continous epidural analgesia with 2% lidoeaine 12 - 14ml. Group Ⅱ ,the patients were injected 0. 6mg/kg katamine into epidural analgesia 30 minutes before operation. Group Ⅲ,the patients were injected 0. 6mg/kg katamine and 1.5μg/kg clonidine into epidural analgesia 30 minutes before operation. The patients in three groups were recorded VAS score on 2h ,4h ,6h, 12h ,24h after operation, also recorded the change of epinephrine(E) and norepinephrine(NE) and sensation and movement recovery time after operation. The side effects such as dizziness nausea,vomit,and exited talking were observed during the operation. Results The VAS score were significantly different between group Ⅰ ,Ⅱ and Ⅲ. The levels of E and NE in plasma in group Ⅰ were increased more than group Ⅱ and Ⅲ within 24 hours after operation, also there is significant difference in group Ⅱ compared with group Ⅲ in T1 ,T2 ,T3. The time of sensation and movement recovery were remarkably longer in group Ⅲ com-pared with group Ⅰ and Ⅱ, showing significant difference. There were no significant difference in side effects after operation in three groups. Conclusion The preemptive analgesia of kutamine and colnidine can relieve the pain of lower abdominal surgery and stress response after operation,and it do not increase the side effects.

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.

Objective To study the dynamic changes and potential impacts on the voice in postoperative pa-tients with different types of thyroid operation.Methods A total of 48 cases thyroidectomy,34 with thyroid isth-mus and lobectomy and 4 with total thyroidectomy,and 19 cases neck operation were included in this study for the examinations of electric laryngoscope,and acoustic voice analysis preoperative and postoperative,respectively.The characteristics of the acoustic parameters were analyzed.Results Vocal cord paralysis and cricothyroid joint disloca-tion in all the patients before and after operation were not observed.For male patients in the experimental group, postoperative parameter values of F0TD were higher than the day before surgery and 30 days after thyroidectomy. The difference was statistically significant (P <0.05 ).In female patients,postoperative parameter values of F0 were lower than the day before surgery and 2nd,7th,and30th day after thyroidectomy.Postoperative parameter values of Jitter for 30th day were lower than 1st day after the operation.The postoperative parameter values of F0TD for 1st,2nd,3rd,and 7th day were higher than the day before thyroidectomy.The parameter value of F0TD was lower than 1st day after surgery,and the postoperative parameter value of HNR for 30th day was lower than 3rd day after the thyroidectomy.The difference was statistically significant (P <0.05 ).Compared to the control group,there were no significantly differences in male patients of experimental group before and after the surgery(P >0.05).The parameter values of F0 in famale patients before the thyroidectomy,the 1st,2nd,3rd,7th day after the surgery and the parameter value of Jitter,Shimmer,HNR for 1st day after the operation were significantly dif-ferent(P <0.05).There were no significant differences in all parameter value of 30 days after thyoidectomy(P >0. 05).Conclusion Voice parameter values alter in a way after thyroid surgery without apparent recurrent laryngeal nerve injury,especially in the female patients.The improvement of voice acoustic parameters can be observed on the 30th day after operation.

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 identify the HLA-A0201 restricted CTL epitopes derived from hepatitis B virus X protein predicted by computer program and general principles in vitro.Methods:HBx gene sequences of Hepatitis B virus genotypes B/C and serotypes adw/adr,with the highest frequencies in Chinese,were computed and analyzed by screening service offered by Internet combined with peptide supermotif,extended motif and quantitative motif prediction.Four most ideal nine-peptides(HBx1,HBx2,HBx3,and HBx4)were selected as candidate peptides.Using flow cytometry,the fluorescence index of both control and experimental groups were detected and the 4 nine-peptides were evaluated with T2 binding assay and DC50 assay.Results:The nine-peptides VLCLRPVGA(HBx1),CLFKDWEEL(HBx2),VLHKRTLGL(HBx3)and HLSLRGLPV(HBx4)were selected as candidate targets.Among the 4 candidate peptides,HBx2 showed higher HLA-A0201 affinity and HBx2,HBx4 showed better stability.Conclusion:Our study indicates that CLFKDWEEL might be a potential HLA-A0201 restricted CTL epitope from hepatitis B virus X protein;further study is needed for verification of its immunity in vivo.

OBJECTIVE To understand the relevant factors of nosocomial infection of inpatients in order to provide the evidence for taking measures to prevent and control the infection effectively.METHODS A retrospective survey was carried out on 10059 cases of hospitalized patients during Jan and Dec of 2008.RESULTS The nosocomial infection rate of hospitalized patients in this period was 4.3%(437/10059).The high risk infected rates were respectively as follows:24.9% in the Blood Department,15.1 % in the Tumor Department;The infection site were respiratory tract(59.3%),urinary tract(14.3%);The main pathogens were Gram negative bacteria(40.7%),fungus infection(25.4%);The risk factors mainly were usage of antibiotic drugs(40.7%),and underlying diseases of tumor(16.2%).CONCLUSIONS The effective measures for reducing the incidence of nosocomial infection should be taken.More attention should be paid on the risk factors.

AIM To study the mechanism of puerarin against arrhythmias. METHODS Standard microelectrode intracellular recording technique was used RESULTS ①Puerarin 0 005, 0 01,0 015 mmol?L -1 prolong the action potential duration at 50% and 90% of repolarization (APD 50 ,APD 90 ) respectively APD 50 was prolonged from (176 43?51 37) ms to (192 86?60 82) ms ( n=7, P