Objective To study the protective effect and Bcl-2 expression of salvia miltiorrhiza pretreatment on retinal ischemia-reperfu-sion injury ( RIRI) . Methods One hundred and thirty two Wistar rats were randomly divided into the normal control group, the ischemia-reperfusion group and the salvia miltiorrhiza pretreatment group. The model of retinal ischemia-reperfusion injury was constructed by increas-ing the intraocular pressure. The ischemia-reperfusion and salvia miltiorrhiza pretreatment group were divided into five subgroups according to the different reperfusion time (6 h, 12 h, 24 h, 48 h and 72 h). Observe the histological changes in retina by HE staining. The SABC ( strept avidin-biotin complex) and Western-blot were used to measure changes of Bcl-2 protein levels in retinal. Results The positive ex-pression of Bcl-2 protein was weak in normal group. In the ischemia-reperfusion group and salvia miltiorrhiza pretreatment group, the expres-sion of Bcl-2 protein began to increase at 6 hours after reperfusion, reached the peak at 24 hours after reperfusion, began to decrease at 48 hours after reperfusion, and started to weaken at 72 hours after reperfusion. The variation tendency of the two groups were the same, however, the expression of Bcl-2 was significantly stronger in the salvia miltiorrhiza pretreatment group compared with ischemia-reperfusion group, and there was significant difference between the two groups (P<0. 01). Conclusion Salvia miltiorrhiza pretreatment can protect the retina by reducing retinal ganglion cells apoptosis in retinal ischemia-reperfusion injury. The mechanism may be achieved by regulating the expression of Bcl-2 protein.

Objective To explore the relationship between preoperative plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) and recurrence of atrial fibrillation (AF) after cardiac valve replacement (CVR) combined with radiofrequency ablation.Methods Fifty-eight patients with AF undergoing CVR combined with radiofrequency modified Maze procedure were enrolled in this study.The patients were allocated to either AF recurrence group (AF group,18 cases) or AF no recurrence group (SR group,40 cases) according to whether patient's AF recurrence or not within 3 months after surgery.NT-proBNP were respectively measured in two groups in 1st day before operation and 9th day after operation by radioimmunoassay.Results All the 58 patients were involved in the final analysis.NT-proBNP levels in 1st day before operation and 9th day after operation in AF group was significantly higher than that in SR group (2061.30 ng/L vs.579.00 ng/L,996.60 ng/L vs.209.20 ng/L,P＜ 0.01 ).NT-proBNP levels in 1st day before operation in two groups was significantly higher than that in 9th day after operation (P ＜ 0.01 ).Conclusions There is a good relationship between preoperative plasma NT-proBNP levels and recurrence of AF.It has a clinical value in predicting of recurrence AF after CVR combined with radiofrequency modified Maze procedure.

Background Acanthamoeba keratitis is a sort of serious infectious eye disease with high causing-blindness rate.Acanthamoeba keratitis often is misdiagnosed as fungal keratitis or viral keratitis in the early stage.Because conventional clinical diagnosis methods show a low specificity and take a long time,timely treatment often is delayed.Conventional PCR does not apply well because the lesion sample is not enough to extract DNA.However,direct PCR can amplify 18S rRNA conserved sequence of acanthamoeba keratitis without the extraction of DNA.Objective This study was to discuss the feasibility for rapid diagnosis of acanthamoeba keratitis using direct PCR to amplify the gene 18S rRNA fragment.Methods Ten acanthamoeba strains were isolated from 10 eyes with acanthamoeba keratitis in Qingdao Eye Hospital.The sensitivity of the direct PCR assay was tested using different numbers of amoebas.The specificity of the assay was tested using DNA extracted from acanthamoeba,candida albicans,pseudomonas aeruginosa,herpes simplex virus-1 (HSV-1) and normal human corneal epithelial cell.Acanthamoeba keratitis models were established using infected method in clean 6-week-old female BALB/c mice.Corneal lesion samples were obtained 1 day,3,5,7,10,15 days after modeled.The effectivity and feasibility of the direct PCR assay for rapid diagnosis of acanthamoeba keratitis were evaluated and compared with culture method,corneal smear examination and real-time PCR.Results Direct PCR primers could only amplify DNA of acanthamoeba rather than other pathogens,and 10 stains of acanthamoeba were detected at least in each sample.During the development of acanthamoeba keratitis in the mice,the diagnosis positive rate of direct PCR was 80.0％,90.0％,80.0％,70.0％,70.0％ and 50.0％ in 1 day,3,5,7,10,15 days after modeled with the total positive rate 73.3％,which was higher than 31.7％ of culture method,56.7％ of corneal smear examination and 61.7％ of realtime PCR,with a significant difference between the direct PCR and culture method (P =0.005),but no significant difference was seen in the total positive rate between the direct PCR and real-time PC R (P =0.172) or corneal smear examination (P =0.056).Conclusions The direct PCR assay is a simple,rapid,highly specific and sensitive method for the rapid diagnosis of acanthamoeba keratitis,especially for the limited lesion sample.

Based upon previous theoretical research, the author puts forward the characteris-tics of excellent teaching team of clinical psychology in universities. According to the situation and the problem of the talents cultivation of clinical psychology in medical university, such as the teachers has lesser skill in clinical psychology, the vague cultivation goal, the faulty curriculum setting and the in-sufficiency practice teaching, the methods improving the teachers' ability and exploring the model and the means of the talents cultivation of the clinical psychology are important during constructing teach-ing team of the clinical psychology.

BackgroundCorneal graft reject is a major cause of corneal transplantation failure.Although many immune-suppressing drugs have been utilized to reduce the reject response,their adverse effects on organ and tissue are still insoluble.The tolerance induction of natural killer T (NKT) cells is currently under investigation.However,the study on the application of NKT cells in high risk corneal transplantation is seldom.ObjectiveThe present study was to explore the effects of α-GalCer-activated NKT cella on allografts survival after high-risk corneal transplantation surgery via retro-bubar injection.Methods The lymphocytes were picked up from the spleen of SPF Lewis rats and cultured in RPMI 1640 medium with 100 mg/L α-GalCer.After one week,NKT cells were sorted by the FACSVantage system as CD161+ TCR-α+ cell from the lymphocytes with the cell densities 5×106/ml.Ten SPF Fisher344 rats were used to prepare the donor corneas,and 20 Lewis rats served as recipients.The high risk corneal transplantation models were created by corneal suturing in 20 recipient rats.Penetrating keratoplasty (PKP) was performed in the model rats.0.1 ml NKT cells or the same volume of normal saline solution were retro-bubarly injected at the end of surgery respectively.The corneal allografts were observed and scored based on Holland criteria at the three-day interval under the slit lamp for 30 days.Two weeks after surgery,three rats from each group were sacrificed by excessive anesthesia method and the eyeballs were obtained for histopathological examination.The inflammatory cell infiltration ( CD4+ and CD8+ ) in grafts was evaluated by immunochemistry and flow cytometry.The use of the animals complied with the Statement of ARVO.ResultsThe mean survival time of the allografts was (7.90± 1.37) days in normal saline solution group and (14.70± 1.49) days in NKT cell group,showing a statistically significant difference between the two groups ( t =10.61,P =0.00 ).Two weeks after surgery,all the allografts showed the severe opacity with lots of new blood vessels and edema in normal saline solution group.However,the corneal grafts were clear in NKT cell group.Abundant CD4+ and CD8+T lymphocytes were seen in the allografts in normal saline solution group,but the inflammatory cells were obviously less in NKT cell group.The percentage of NKT cells in the spleen was (5.67±0.25)％ in NKT cell group and ( 1.21±0.19)％ in normal saline solution group ( t =8.43,P =0.00 ).Conclusionsα-GalCer-activated NKT cells can prolong the survival time of allografts in high-risk corneal transplantation.Retro-bubar injection of α-GalCer-activated NKT cells probably is a new approach to the prevention of the rejection of corneal transplantation.

Objective To evaluate the utility of susceptibility weighted imaging (SWI)in the evaluation of traumatic brain injury (TBI)severities.Methods 20 mild TBI patients,20 moderate TBI patients and 20 severe TBI patients were collected.The involving brain regions,numbers and areas of hemorrhagic lesions detected by SWI were analyzed and the differences of each group were com-pared.The correlation analysis between Glasgow coma scale(GCS)scores and each above parameters were underwent.Results The differences of involving regions’number,lesions’number,lesions’area were statistically significant(P <0.05 ).Severe TBI group got the maximum number of involving regions,lesions,and the largest areas,followed by moderate group and mild group.The GCS were highly negatively correlated with the number of involving regions,number of lesions,areas of lesions detected by SWI.In descending order of relevance:GCS with areas detected by SWI(r=-0.982,P=0.000),the number of lesions detected by SWI(r=-0.941,P=0.000),the number of involving regions detected by SWI(r=-0.900,P=0.000).Conclusion The clinical applications of SWI in diagnosing TBI and evalua-ting the TBI severity is of great significance.

Objective:To investigate the inhibitory effect of Dickkopf-1 (Dkk1) on vasculogenic mimicry (VM) formation and the relevant mechanism. Methods:CD34-PAS dual staining and immunohistochemical staining were used to detect and analyze the re-lationship between VM existence and Dkk1 expression in 217 human colon cancer tissue samples;three dimensional (3D) culture was used to detect the influence of Dkk1 on tube structure formation and on VE-cadherin expression;a subcutaneous mouse xenograft mod-el was made to further validate the inhibitory role of Dkk1 on VM formation in vivo. Results:VM-positive samples indicated a lower expression of Dkk1(P<0.05);colon cancer cells with Dkk1 overexpression exhibited a decreased ability to form tube-like structure and a decreased expression of VE-cadherin;Dkk1 inhibited the VM-formation abilities of human colorectal carcinoma cell line xenograft tu-mor tissue. Conclusion:Dkk1 inhibits the VM formation of colon cancer.

Objective To evalute the accuracy of stroke volume variation (SVV) in monitoring blood volume in patients undergoing off-pump coronary artery bypass grafting.Methods Twenty-one ASA Ⅱ or Ⅲ patients of both sexes aged 44-77 yr undergoing off-pump coronary artery bypass grafting were enrolled in this study.Anesthesia was induced with midazolam,etomidate,fentanyl,rocuronium and dolicaine and maintained with target-controlled infusion of propofol,infusion of remifentanil,intermittent iv injetion of atracurium and inhalation of sevoflurane.The patients were mechanically ventilated (VT 8 ml/kg,RR 12 bpm,I:E 1:2,PEEP 0,FiO2 80％ ).PEr CO2 was maintained at 35-44 mm Hg.Radial artery was cannulated and connected to FloTrac pressure transducer and Vigileo monitor.6％ hydroxyethyl starch 130/0.4 sodium chloride solution 7 ml/kg was infused at a rate of 0.25 ml· kg- 1 1· min- 1 at 5 min of haemodynamics stabilization after pericardiotomy (T1).HR,MAP,CVP,systemic vascular resistance (SVR),systemic vascular nesistance index (SVRI),SVV,stroke volume index (SVI)and CI were recorded at T1 and at 10 min after loading dose (T2).The change rate of HR(△HR),MAP(△MAP),CVP(△CVP),SVR(△SVR),SVV(△SVV),SVI(△SVI) and CI(△CI) were calculated.△SVI≥25％ was considered effective volume expansion.The ROC curves for HR,MAP,CVP,SVR and SVV in determining the volume expansion efficacy were plotted.The area under the curves and 95 ％ confidence interval were calculated.Results Compared with T1,CVP,SVI,CO and CI were significantly increased,SVRI and SVV decreased at T2 (P ＜ 0.01).There was no significant difference in MAP and HR between T1 and T2(P ＞0.05).△SVI was negatively correlated with △HR and △SVR ( r =- 0.737,r =- 0.480,P ＜ 0.05).△SVI was not correlated with △CVP,△MAP and( P ＞ 0.05).The change in SVI was determined by SVV 8.8％ (sensitivity =52.6％,specificity =100.0％ ).The area under the curve for SVV and 95％ confidence interval were 0.579(0.346-0.812).Conclusion SVV can not be used to accuratelymonitor the changes in blood volume in patients undergoing off-pump coronary artery bypass grafting.

Objective To explore the effect of high voltage alternating electric field therapy on fracture healing in rats. Methods An animal model of cortical bone defect was successfully established in the unilateral proximal tibia of 41 rats.They were then randomized into an experimental group (21 rats) and a control group (20 rats).The animals in the experimentsl group were given high voltage alternating electric field therapy for 30 minutes daily,while those in the control group only had sham exposure to the high voltage alternating electric field therapy.Rats in each group were sacrificed at days 7,14,21 and 28 post-operation.After blood collection,the operated tibias of the rats were resected with muscle and other soft tissue removed.Bone mineral density (BMD) of the callus at the fracture point was measured using dual energy X-ray absorptiometry (DEXA).The calluses were then cut down and fixed,decalcificated and sliced for histological observation.At the same time serum osteocalcin ( OC ) and bone alkaline phosphatase (RAP) were also detected by ABC-ELISA. Results The BMD examination showed that callus BMD in the experimental group was significantly higher than in the control group at days 14 and 28.Serum OC and RAP in the experimental group were also higher at days 14 day 21,and OC was still significantly higher in the experimental group at day 28.Histological examination showed that in the early stages of bone healing hematoma absorption and organization appeared earlier in the experimental group than in the control group,with remarkable osteoblast and chondroblast proliferation.While at the later stage,bone trabeculas connection and mineralization also appeared earlier in the experimental group than in the control group. Conclusions High voltage alternating electric field therapy may promote fracture healing by accelerating hematoma absorption and organization,elevating osteoblast proliferation and differentiation as well as promoting extracellular matrix secretion and mineralization.The underlying therapeutic mechanism might be related to the increased expression of OC and RAP caused by exposure to the high voltage alternating electric field.

Objective To look for a reliable and convenient judgement criteria for the screening of cytomegalovirus pneumonia in order to reduce misdiagnosis and resulted mistherapy.Methods Process collected data on fifty-six cytomegalovirus pneumonia and forty-two common viruses induced asthmatic bronchitis cases by use of discriminant analysis to construct prediction model of diagnosis result.Results Only three indexes including age,lymph count and platelet count were selected into the model via sift.The performance of the established screening model showed as follows:sensitivity was 80.36％,specificity was 80.95％,misdiagnosis rate was 19.05％,false negative rate was 19.64％,diagnostic accordance rate was 80.61％.Conclusion Being concise and of strong maneuverability and high accuracy in prediction,cytomegalovirus pneumonia diagnosis model constructed through discriminant analysis can provide powerful screening means for medical staff.