Aim To investigate the protection of ramipril,BQ-123 and their combination against myocardial ischemia/reperfusion(I/R)injury in vivo in anesthetized rats,and to explore the mechanism of action of drugs on myocardial oxidation-antioxidation system.Methods Healthy male Wistar rats were divided into 5 groups randomly,sham operated(Sham)group,I/R group,ramipril(RAM)group,BQ-123(BQ)group and ramipril and BQ-123(R&B)group.All groups but not sham were subjected to I/R procedure.Twenty four hours before ligation,ramipril(1 mg·kg~(-1))was intragastrically administered to rats in RAM and R&B groups.The equal volume of normal saline was given to rats in other groups.BQ-123(10 μg·kg~(-1)· min-1)was infused intravenously from 10 min before ligation to the end of 30 min ischemia to rats in BQ and R&B groups.The equal volume of normal saline was given to other groups.HR,MAP and the change of ST-segment were observed;ventricular arrhythmias were monitored during ischemia;the infarct size was examined by TTC staining;the activity of myocardial T-SOD,Mn-SOD,CAT and the content of MDA were detected by spectrophotometer.Results Compared with I/R group,the elevation of ST-segment was decreased,onset of VPC and VT was delayed,duration of VPC and VT was shortened,incidence of VPC,VT and VF was decreased,IS and IS/AAR were improved,activity of T-SOD,Mn-SOD and CAT was increased,the content of MDA was decreased in RAM,BQ and R&B groups.Compared with RAM and BQ alone group,onset of VPC and VT,duration of VPC and VT,size,activity of T-SOD and Mn-SOD and content of MDA were changed dramatically in R&B group.Conclusions Ramipril,BQ-123 and the combined use of these two agents protected myocardium from I/R injury in vivo.The protective effects of the combination on delaying onset of VA,shortening duration of VA,decreasing infarct size and content of MDA,and increasing activity of SOD are better than those of using ramipril or BQ-123 alone.

Objective To investigate the changes of circulaling progenitor cells and endothelial progenitor cells(EPCs)in non-septic and septic shock patients using flow cytometry.Method A total of 27 sepsis patients hospitalized in emergency department of Zhongshan hospital during August 2007 to February 2008 were enrolled in this study.The patients were dividedinto septic shock group(n=12)and non-septic shock group(n=15).Ten healthy individuals and ten non-sepsis ICU patients were collected as controls.Peripheral blood mononuclear cells(PBMCs) were isolated by Ficoll density gradient centrifugation,and EPCs labelled with antibodies against CDl33,CD34 and VEGFR-2 were identified and isolated by three-color fluorescence flow cytometry.Differences within the groups were analyzed using One way ANOVA.Results The percentages ofprogenitor cells and EPCs in the PBMC fraction in healthy controls were(0.25%4-0.14%),(0.09%-I-0.02%),respectively,and those in ICU controls were(O.38%.4-0.29%),(0.12%.4-0.02%).The percentages of progenitor cells and EPCs were significantly higher in栅sel如c shock patients(0.57%±0.12%),(0.22%.4-0.10%)than in heathy and non-sepsis ICU controls(P<0.05).However.the percentages of progenitor cells and EPC8 in septic shock pa.tienta(O.20%±0.12%,0.04%-t-O.01%)was obviousely lower than those in healthy,ICU controls and ilionseptic shock patients(P<0.05).Sptic shock survivors had significantly higher numbers of cEPCs than nonsur.vivors(P<0.05).Conclusions The level of progenitor cells and EPC8 in peipheral blood of sepsis patients might be the valuable markers to as.se88 the severity and outcome ofthese ptienS.

ObjectiveTo explore relationship between psychological resilience and life events of Tibetan nursing students receiving free vocational nursing education.Method The questionnaire of Symptom Checklist 90(SCL-90)and ASLEC were used in the survey among 120 nursing students to investigate the psychological resilience and life events and SPSS 17.0 was used to look into the correlations between them.Results The scores on psychological resilience of Tibetan nursing students was higher than the norm (P<0.05).There were negative correlations between life events and psychological resilience(P<0.05).Their psychological resilience was positively related to personal relations and health conditions(P<0.05)but negatively to learning pressure,punishment and losses (P<0.05).Conclusion The school and hospital should make teaching measures to psychologically support them in daily life to strengthen the abilities to handle failure and adjust to the society.

Objective To evaluate the value of plain and dual-phasic enhanced 16-slice CT in the diagnosis and preoperative TNM staging of the gastric carcinoma,and to discuss the relationship between image signs and pathologic findings.Methods Fifty-three cases of the gastric carcinoma confirmed histopathologically underwent 16-slice CT examination.The scan protocol included plain scanning,the arterial phase and portal venous phase scanning.The manifestation of the three series images and multiplanar reconstruction(MPR) images were analyzed.Results ①The accuracies of 16-slice CT for the T stage,the N stage and the M stage of the gastric carcinoma were 83.02%,80.00% and 92.45% respectively.②The overall accuracy of 16-slice CT for judging TNM stage was 84.91%.Conclusion The plain scan and dual phase enhanced scans of 16-slice CT,especially the thin slice and MPR with proper windows technique are helpful for the diagnosis of gastric carcinoma and the TNM stage,which is useful for the selection of the operative project and the therapy plan.

OBJECTIVE:To establish HPLC method for the simultaneous content determination of metronidazole,tetra-caine hydrochloride and dexamethasone acetate in metronidazole compound.METHODS:The content determination was per-formed on column C 18 ;the mobile phase consisted methanol-water-triethylamine(65∶35∶0.36),the detection wavelength of both metronidazole and tetracaine hydrochloride was310nm,and240nm for dexamethasone acetate,the flow rate was1ml/min and the column temperature was25℃.RESULTS:The linear ranges of metronidazole,tetracaine hydrochloride and dexameth_ asone acetate were0.412?g～2.06?g(r=0.9999),0.191?g～0.956?g(r=0.9999)and0.121?g～0.604?g(r=0.9998),respectively,and their average recoveries were100.69%(RSD=1.28%),101.37%(RSD=0.23%)and102.40%(RSD=0.89%),respectively.CONCLUSION:The method was simple,accurate,reproducible,and suitable for the quality control of metronidazole compound.

Objective To investigate the features of gallbladder carcinoma in two-phase spiral CT, and to analysis the values of two-phase spiral CT for the differential diagnosis between gallbladder carcinoma and chronic cholecystitis. Methods The two-phase spiral CT manifestations of 30 cases of gallbladder carcinoma, proved by surgery and pathology, and 30 cases of chronic cholecystitis were analyzed. Results According to the CT findings, the gallbladder carcinoma was categorized into 3 types: intraluminal mass of gallbladder in 6 out of 30 (20.0%), thickening of the gallbladder wall in 11 (33.7%), and mass replacing the normal gallbladder in 13(43.4%). The most common enhancement patterns of the wall in gallbladder carcinoma were hyperattenuation during the arterial phase, while isoattenuation with the adjacent hepatic parenchyma during the venous phase; or hyperattenuation during both phases. The most common enhancement pattern of the wall in chronic cholecystitis was isoattenuation during both phases, with clear hypoattenuation linear shadow in the gallbladder fossa. Other ancillary features of gallbladder carcinomas included: infiltration of the adjacent parenchyma, local lymphadenopathy and intrahepatic metastasis. Conclusion Two-phase spiral CT scan can identify the features of the gallbladder carcinoma and is helpful for the differential diagnosis of these two different disease entities.

Objective To study the effect of ischemia reperfusion(IR) on electrophysiological function of sinoatrial node(SAN) in rabbits. Methods Ninety healthy adult rabbits were divided randomly into control group, ischemia groups (I) 10(I10 min), 30(I30 min), 60(I60 min) and 120 min(I120 min) and IR groups (10, 30, 60 and 120 min ischemia followed by 4 h reperfusion respectively)(I10 minR4 h, I30 minR4 h, I60 minR4 h and I120 minR4 h). There were 10 rabbits in the control group and each subgroup. IR injury model of SAN was established by occluding and loosening the start section of right coronary artery(RCA). The changes of AA interval and arrhythmia were recorded by electrocardiography and cardiac chamber electrography synchronously. Results ① There was no significant difference in AA interval between different time points in the control group. ② Sinus arrest(SA), sinus bradycardia with arrhythmia(SB), or atrial tachyarrhythmia(AT) were found in 51 rabbits (63.75%) out of 80 rabbits in I and IR groups in ischemic period. The incidence rate of arrhythmia increased in a time dependent manner. The AA intervals also increased by at least 40 ms in 54 rabbits (67.5%). ③ Sinus or atrial arrhythmia during ischemic period was found in 26 out of the 40 rabbits in IR group, but 15 returned normal after reperfusion. ④ Increased AA intervals were found in 27 out of the 40 rabbits in IR group during ischemic period. Most of them recovered to pre occlusion level within 10 min after reperfusion, but the AA intervals prolonged again in I60 minR4 h and I120 minR4 h groups as the reperfusion elongated. Conclusion ① These findings suggest that about 2/3 of the rabbit sinus node arteries may stem from right coronary artery. ② Electrophysiological changes due to ischemia of SAN resemble the electrocardiogram of sick sinoatrial node syndrome. ③ Reperfusion arrhythmia can be induced by reperfusion after a long time of ischemia.

BACKGROUND:There is a linker between Hsp65 and Esat6 coding a segment of water repellent polypeptide.It is very gentle and easy to be folded,which is profitable to translate the keno-folding correctly between the two proteins and to make the space structure of fusional proteins consistent with those two native ones.Then,a correct structure is formed and the immunogenicity of the fusional proteins is improved.OBJECTIVE:This study was to clone the fusional genes Hsp65-Esat6 from mycobacterium tuberculosis(MTB)H37Rv, then to construct into a eukaryotic expressing vector which contains an enhanced green fluorescent protein(EGFP) reporter gene,and finally to identify the expression of Hsp65 protein and Esat6 protein by IHC methods.DESIGN:Single sample experiment.SETTING:Department of Microbiology ＆ Immunology,Medical College of Jinan University.MATERIALS:Plasmid pVAE was donated by Professor Cao from South China University of Technology.MTB H37Rv,E.coil DH5 α,expressing plasmid pEGFP-C1 and Hela cells were donated by Doctor Hu Ping.METHODS:The experiment was conducted at the Department of Microbiology ＆ Immunology and the Medical Experimental Center of Jinan University between September 2005 and June 2006.The whole-genome was extracted from MTB H37Rv by molecular cloning technique,and used it as template to amplify Hsp65 (no terminator) gene by polyrnerase chain reaction (PCR),to recombine it with pEGFP-C1 vector after purification to construct pEGHsp65(no terminator)recombination vector.pVAE vector was used as template to amplify Linker-Esat6 gene(with terminator)by PCR, and then to recombine it with pEGHSP65(no terminator)to construct an eukaryotic expressing vector pEGHsp65-Esat6 with the fusional genes Hsp65-Esat6 inside.Finally,genes Hsp65-Esat was checked by molecule biology methods such as PCR, restriction endonuclease and DNA sequencing.Hela cells were transfected with pEGHsp65-Esat6 and the expression of EGFP and the efficiency of transfection were observed.The expressions of Hsp65 protein and Esat6 protein were detected by IHC methods.MAIN OUTCOME MEASURES:①The size of pEGHsp65-Esat6.②The DNA sequencing result of the pEGFP-C1.③The expression of EGFP in the transfected Hela cells.④The IHC results of Hsp65 protein and Esat6 protein in Hela cells.RESULTS:①The size of pEGFP-C1 was 4.7 kb,that of pEGHsp65 was 6.4 kb,and that of pEGHsp65-Esat6 was 6.7 kb.There were differences between their speeds In agarose electrophoresis.②The results showed that it was the same as reported Hsp65 sequence and Esat6 sequence of MTB H37Rv.③After transfecting pEGHsp65-Esat6 for 24 hours,EGFP was found in 30% of Hela through Laser scanning confocal microscope. But there was no EGFP in non-transfected Hela.④Hsp65 protein and Esat6 protein with biological activities were detected in transfected Hela cells by IHC methods.CONCLUSION: Using Hsp65 and Esat6 as immunogens,we have successfully cloned and constructed a eukaryotic expressing vector which contain fusionaI genes Hsp65-Esat6 of MTB H37Rv and EGFP.

Objective To explore the endoscopic managements for biliary and pancreatic diseases in patients with the history of complex gastrointestinal surgery.Methods Data of four patients who underwent balloon-assisted enteroscopy after complex digestive surgeries (2 patients underwent Roux-en-Y reconstruction,2 others Whipple reconstruction) were retrospectively collected.One patient with bilio-intestinal anastomosis was explored by double-balloon enteroscope (DBE),and balloon-assisted enteroscopy combined with ERCP was used for other 3 patients.Results Therapeutic ERCP was successfully performed on one patient,and the diagnosis of 3 others were confirmed.No operation-related complications such as bleeding or perforation was observed.Conclusion Balloon-assisted enteroscopy combined with ERCP is an important diagnostic and therapeutic method for biliary and pancreatic diseases in the patients after complex gastrointestinal surgery.

Objective To develop a novel method for treating complete heart block by autotransplantation of simus node node cells to right ventricular anterior wall.Methods Twenty healthy mongrel dogs were involved in the present study.The dogs were randomly assigned to transplant group or control group(n=10).The sinus node (SN)was harvested and isolated in vitro after an electronic pacemaker was implanted and complete heart bolck was introduced.The SN cells from dogs of transplant group were injected to autogenic right ventricular wall.Commensurable culture medium was implanted to ghe same position of dogs in control group.Two weeks later,detailed electropohysiological study was performed.For investigating the variation of the rhythm,epinephrine was administrated through femoral vein to dogs of transplant group.Results Most of isolated SN cells from dogs were thin-spindle shape,and cell activity was fine.The SNs by VG stained displayed typical structural feature.2 weeks after cell autotransplantation,higher heart rates were achieved from transplant group than that in control group(P＜0.05).This rhythm was stable in 4 weeks and became faster remarkably after administration of eninephrine(P＜0.05).Conclusion SN cell of dogs tutorgrfted into right ventricular anterior wall can form new pacemaker site in ventrcle and improve ventricular rate of complete heart bolck.This pacemaker site can also be regulated by epinephrine.