Purpose　To study the proteins expression of genes related to apoptosis of retinal cells in development of human fetus．　Methods　Fifty cases of retinas of human fetus aged from 12 to 38 weeks were collected and paraffin embedded sections were made． Immunohistochemical method was used．　Results　Fas protein was expressed by cells of ganglion cell layer, inner and outer nuclear later, which were just formed on 16th week． It was not expressed until 38th week, Fas(+) staining appeared in layers of retina． Fas-L(+) staining was detected in cells of layers of retina on 26th week and the positive staining located in ganglion cell layer on 32th week． Neuronal fiber layer was Fas-L positive． Bax positive staining was detected on 8th week． Bax positive nucleus were observed mainly in GCL and ONL on 16th week． It was in INL on 24th week and in Müller cells inner terminates on 26th week． After this time, all cells of retina were bax immune negative staining． Bcl-2(+) staining appeared in differentiating neuroblastic layer on 16th week． Beginning on 24th week, bcl-2 (+) staining was observed in glial cells of GCL and inner terminates of Müller cell．　Conclusion　Apoptosis of developing retinal cell may be Fas/Fas-L independent and bax may be involved in apoptosis of the cells．

ObjectiveTo observe the effect and adverse reaction of Reduning injection in the treatment of epidemic hemorrhagic fever.Methods98 patients with epidemic hemorrhagic fever were randonly divided into 2 groups,the treatment group was given Reduning injection ;The control group was given ribavirin injection;The efficacy and prognosis and adverse reaction were compared between two groups.ResultsThe antipyretic effect of the treatment group was better than that of the control group ( x2 =6.41,P ＜ 0.05 ),and the fever clearance time,oliguria time,the total duration was significantly shortened( t =2.356,2.071,5.125,all P ＜ 0.05 ).There were no obvious adverse reaction in the two groups.ConclusionReduning injection in the treatment of epidemic hemorrhagic fever had good curative effect,safety,and it was worthy of popularization and application.

Objective To investigate the clinical effect of aspirin and ursodeoxycholic acid on treating intrahepatic cholestasis of pregnancy (ICP).Methods Seventy-one ICP patients were divided by random digits table method into observation group (36 cases) and contol group (35 cases).Observation group was treated with aspirin and ursodeoxycholic acid.Control group was given unsodeoxycholic acid.The biochemical indicators,itching score and pregnancy outcomes of two groups were observed.Results Total bile acid (TBA),total bilirubin (TBIL),alanine aminotransferase(ALT),aspartate aminotransferase(AST)and itching score of two groups after treatment were all significantly lower than those before treatment (P ＜0.05 ).TBA,TBIL,ALT and itching score of observation group[ (31.7 ± 8.3 ) μ mol/L,(33.9 ± 5.8 ) μ mol/L,(53.1 ± 8.4) U/L,(0.48 ± 0.06) scores] were obviously lower than those of control group [(76.2 ±9.2)μ mol/L,(56.9 ± 8.2) μmol/L,(80.8 ± 10.6) U/L,(0.81 ±0.04) scores] (P ＜0.05).The incidences of amniotic fluid contamination,premature delivery and neonatal asphyxia of observation group were significantly lower than those of control group [38.9％(14/36) vs.71.4％(25/35),11.1％(4/36) vs.31.4％( 11/35 ),5.6％ ( 2/36 ) vs.22.9％ ( 8/35 ),P ＜ 0.05 ].Conclusions The combined application of aspirin and ursodeoxycholic acid can improve the biochemical indicators,itching symptom and pregnancy outcomes.

The paper reviewed clinical reports on the treatment of hypermenorrhea, menostaxis, and uterine bleeding with Ancong Decoction and modified Ancong Decoction. Although there were few reports concerned with clinical usage, Ancong Decoction has sound therapeutic effects, with more than 90% effective rate.

Objective:To prepare the vaccine of DC derived from human peripheral blood and transfected with HPV16E6 antigen gene, and to detect its morphological character,surface marker and immunological effect.Methods:DC-enriched populations were prepared from human peripheral blood mononuclear cell(PBMC) with the combination of rhGM-CSF,rhIL-4 and rhTNF-?. The plasmid containing HPV16E6 gene was transfected into DC with lipofectamine. The morphology of DC was observed dynamically, and the expression of surface markers of DC vaccine could be detected using immuno-cytochemical staining and flow cytometry. MTT assay was applied to detect the activity of CTL in vitro.Results:The transfected DC had typical morphologic and phenotypic characteristics, and expressed E6 protein 47.3%, CD80 82.5%, CD86 79.8% and CD83 85.7%. The killing activities of CTL to Caski cells induced by transfected DC were higher evidently than that of control groups(P

BACKGROUND: Two-dimensional electrophoresis (2-DE) is precise and effective for the isolation and analysis of complex protein mixtures, has become one of valuable key techniques for proteome. Mice models were as research subjects to model human disease status, one of important methods for cardiovascular disease. The establishment and optimization of 2-DE for proteomes of mice myocardium will provide basis for heart disease.OBJECTIVE: To establish and optimize a stable technique of 2-DE for proteomic analysis of mice myocardium.DESIGN: Mice were used as research subjects, observational comparative study.SETTING: Department of Pathology, Nanfang Hospital Affiliated to Southern Medical University.PARTICIPANTS: The experiment was performed at the Department of Pathology, Nanfang Hospital Affiliated to Southern Medical University between February and September 2004. Totally 12 male BABL/c mice of 4-5 weeks without special-pathogen free with the body mass of 12-15 g were selected, which was provided by the Experimental Animal Center of Southern Medical University.METHODS: The mice were put to death by dislocation of cervical vertebra under drugged state to obtain heart. Various protein extraction method,loading sampling buffer, strip range of IPG, sampling volume of protein,staining method and so on were compared, analyzed, scanned and digilized,and then performing image analysis. Each link in the study was established and optimized.MAIN OUTCOME MEASURES: Resolution of protein and reproducibility of the trial.RESULTS: A method that was optimal for cardiac tissue can isolate about a total of 910 protein spots on pH 4-7 gel strips. 200 μg was fit for silver stain while 1000 μg was for Coomassie stains. It was found that in the myocardium of normal mice about (920±30) protein spots were obtained in 17 em pH 3-10 L gel strip, while around (880±30) were gained in the pH 4-7 L gel strip. The mean matching rate achieves to 86. 9%.CONCLUSION: The findings of the trial showed that 2-DE technique can be effectively applied for proteomics of myocardium of mice to regulate and optimize, and relatively stable separation method of myocardial protein was established.

Objective To study the thrombolysis effect and safety of UK combined with tirofiban in young patients with ST segement elevation acute myocardial infarction.Methods 76 young patients with ST segement elva-tion acute myocardial infarction were selected as research objects,and they were divided into UK group(control group, n =34)and UK combined with tirofiban group(observation group,n =42)by random number table,then the recanali-zation rate of infarct -related artery and incidence of complications were compared.Results The recanalization rate of infarct -related artery in the observation group was 97.6%,which was 91.2% in the control group,there was no significant difference between the two groups(χ2 =1.564,P >0.05);The vascular infarction related to blood flowⅡlevel of the patients was 90.5% in the observation group,which was significantly higher than 70.6% in the control group(χ2 =4.945,P <0.05);The death,complications incidences had not significant differences between the two groups(χ2 =1.252,2.837,2.837,all P >0.05).Conclusion The thrombolysis effect of UK has a high recanaliza-tion rate of infarct -related artery in young patients with ST segement elevation acute myocardial infarction,and the thrombolysis effect of UK combined with tirofiban is better,and it does not increase the incidence of complications.

Objective To observe the expression of p53, bcl 2 genes, vascular endothelial cell growth factor (VEGF), basic fibroblast growth factor (bFGF), insulin like growth factor I (IGF I), and the receptors of these factors of retinal vascular endothelial cells (VECs) of 1 to 20 week diabetic rats, and the relationship between the expressions and cell cycle arrest. Methods Retinal sections of diabetic rats induced by alloxan were immunohistochemically stained and observed by light microscopy (LM) and electron microscopy (EM). Dot blotting and Western blotting were used to determine the expression of mRNA, proteins of p53 and bcl 2. Results Under LM, immunohistochemical positive expression of p53 and bcl 2 were found on the vessels of ganglion cell layer and inner nuclear layer of retinae of 8 to 20 week diabetic rats; under EM, these substances were observed depositing in VECs. The retinal VECs also expressed VEGF, bFGF, IGF I and their receptors. There was no positive expression of other cell types in these retinae, all cell types of retinae in control group, or all cells of retinae of diabetic rats with the course of disease of 1 to 6 weeks. The result of dot blotting revealed that retinal tissue of 20 week diabetic rat expressed p53 and bcl 2 mRNA, and the result of Western blotting revealed that they also expressed p53 and bcl 2 proteins. But retinal tissues of control group did not. Positive expression of bax was not found in the retinae in control group or 1 to 20 week diabetic rats. Conclusion p53, bcl 2 may introduce cell cycle arrest of VECs of retinae in 8 to 20 week diabetic rats. High glucose might stimulate the expression of VEGF, bFGF, IGF I and their receptors, and the growth factors may keep VECs surviving by self secretion.

Objective To investigate the changes in the expression of macrophage migration inhibitory factor (MIF) mRNA in a rat model of ventilator-induced lung injury.Methods Thirty adult male Sprague-Dawley rats,weighing 2β5-260 g,were randomly divided into 3 groups (n =10 each) using a random number table:control group (group C),small tidal volume (VT) mechanical ventilation group (group S) and large tidal volume mechanical ventilation group (group L).The animals were anesthetized with intraperitoneal ketamine 100 mg/kg,midazolam 0.2 mg/kg and atropine 1.0 mg/kg.The rats were tracheostomized and spontaneous breathing was maintained in group C,while the rats were tracheostomized and mechanically ventilated for 4 h in groups S and L.The tidal volume was 7 ml/kg (group S) or 40 ml/kg (group L),I ∶ E was 1 ∶ 1,RR was 80 bpm and FiO2 was 100％.At 4 h of spontaneous breathing or mechanical ventilation,broncho-alveolar lung lavage fluid (BALF) was collected for determination of the total protein concentration,white blood cell (WBC) counts and concentrations of MIF,IL-6 and IL-1β (by ELISA).Then the rats were sacrificed and the lungs removed for microscopic examination and for determination of wet to dry lung weight ratio (W/D ratio) and expression of MIF mRNA (by RT-PCR).Results Compared with C and S groups,WBC counts,concentrations of total protein,MIF,IL-6 and IL-1β in BALF,and W/D ratio and expression of MIF mRNA in lung tissues were significantly increased in group L (P ＜ 0.05).There was no significant difference in the indexes mentioned above between group C and group S (P ＞ 0.05).The pathological changes occurred in group L.Conclusion The up-regulation of MIF mRNA expression in lung tissues may be involved in the development of ventilator-induced lung injury in rats.

Objective To investigate the role of Toll-like receptor9 (TLR9)-myeloid differentiation factor 88 (MyD88) signal pathway in alveolar macrophages in ventilator-induced lung injury (VILI).Methods 30 adult male Sprague-Dawley (SD) rats were randomly assigned to three groups (with 10 rats in each group).Group A was the control group,with spontaneous respiration after tracheostomy.Rats in group B received mechanical ventilation for 4 hours with normal tidal volume (VT) 7 ml/kg after tracheostomy,and group C rats received mechanical ventilation with VT 40 ml/kg for 4 hours.After termination of ventilation,examination with transmission electron microscopy was performed to observe the ultrastructure changes in alveolar epithelial cell type Ⅱ (AEC Ⅱ) of the lung.Lung wet/dry ratios (W/D) and total protein concentration,the concentration of interleukins (IL-6 and IL-1 β) in bronchoalveolar lavage fluid (BALF) were determined.The protein and mRNA expressions of TLR9,MyD88 and nuclear factor-κB (NF-κB) in alveolar macrophages were assayed by Western Blot and real-time reverse transcription-polymerase chain reaction (RT-PCR).Results The ultrastructure of AEC Ⅱ in the group A and group B was almost normal,whereas the chromatin of the nuclei,the lamellar corpuscles in the cytoplasm,the cell membrane and the microvilli of the AEC Ⅱ in the group C showed injurious changes in various degrees.When the group C was compared with the group A and the group B,it was shown that the W/D ratios (5.54 ± 0.17 vs.4.58 ± 0.17,4.69 ± 0.16) and total protein concentration (g/L:6.33 ± 0.61 vs.0.45 ± 0.05,0.47 ± 0.04),IL-6 (μg/L:1.989 ± 0.103 vs.1.033 ± 0.061,1.010 ± 0.069) and IL-lβ (ng/L:2.79 ±0.25 vs.1.05 ±0.15,1.23 ±0.22) in BALF,the protein expressions of TLR9,MyD88 and NF-κB [TLR9 (A value):0.770 ±0.042 vs.0.300 ±0.027,0.310 ±0.037; MyD88 (A value):0.950 ±0.091 vs.0.560 ±0.082,0.580±0.084; NF-κB(A value):1.020 ±0.076 vs.0.740 ±0.052,0.700 ±0.076] in alveolar macrophages were all increased significantly,and all of which showed significant difference (P＜0.05 or P＜0.01).The mRNA levels of TLR9,MyD88 and NF-κB in alveolar macrophages in the group B were (1.13 ± 0.32),(1.18 ± 0.33),and (1.11 ± 0.22) folds of those of the group A,respectively,but there were no significant differences (all P＞0.05).While the mRNA levels of TLR9,MyD88 and NF-κB of alveolar macrophages in the group C were (8.66 ± 0.69),(6.41 ± 0.53) and (5.29 ± 0.71) folds of those of the group A,respectively,and all of them showed significant difference (all P＜0.01).Conclusion TLR9-MyD88 signaling in alveolar macrophages plays a role in pathogenesis of VILI.