Objective:To investigate the immunogenicity of pORF5 plasmid protein,and further to screen and identify its im-munodominant domian.Methods: 10 different fragments of pORF5 gene including full length were amplified from the DNA of Chlamydia trachomatis serovar D by PCR and cloned into appropriate site of pGEX-6p vector to construct recombinant vectors after digested with BamHⅠand NotⅠrestriction endonucleases.After identification by PCR and sequencing,the recombinant plasmids were transformed into XL1 Blue E.coli to express the GST fusion proteins.ELISA and Western blot were carried out to identify the immunogenicity and immunoreaction of pORF5 plasmid protein.10 different fragments were reacted with sera from patients urogenitally infected with Chlamydia trachomatis, mouse polyclonal antibodies and mouse monoclonal antibodies of pORF5 plasmid protein with ELISA method.Results: pORF5 plasmid protein displayed strong immunogenicity and could induce a strong antibody response in human.The reactivity of human antibodies almost completely disappeared,when the native structure of pORF5 plasmid protein was de-stroyed.F6 that only lacked the N-terminal 66 amino acids was recognized by antibodies in ELISA as strongly as the whole pORF5 plasmid protein was.However,no other fragments were significantly recognized although there was a minimal reactivity of F2 and F3 with antibodies.Conclusion:pORF5 plasmid protein was an immunodominant antigen containing conformation-dependent epitope,and the C-terminal three quarters of pORF5 amino acid sequence was required for maintaining its immune dominance and conformation.The significance of the above findings lay a foundation for the further study on pORF5 protein function and vaccine development.

Pulmonary vein isolation (PVI) is a new ,effective and radical method to cure atrial fibrillation .Within a period after radiofrequency ablation (RFA) ,coagulation system is activated in patients ,then thrombus incident such as cerebral embolism may happen .The present article made a review on its mechanism .

Objective To investigate the effect of aldehyde-free fixation solution and aldehyde fixation solution on DNA and cytoplasm of nucleus.Methods The DNA quantification (IOD),discrete coefficient (CV),DNA index (DI) and nucleus area (area) were measured by DNA quantification system (ICM) in 8 different fixation solution;combined with cytological staining,the effects of different fixative solution on nuclear DNA and cytoplasmic staining were analyzed.Results Aldehyde fixation solution was better than aldehyde free fixation solution,the formaldehyde in fixtion solution has important influence on Feulgen-eosin staining.Conclusion Aldehyde fixation solution combined with cytological staining can obtained good dyeing effect,which provides the basis for the subsequent multi-technique joint diagnosis.

Objective To investigate the efficacy of combined application of laparoscope and multiple endoscopes for the treatment of extra-hepatic bile duct stones.Methods The clinical data of 3780 patients with extra-hepatic bile duct stones who were admitted to the First Hospital of Lanzhou University from March 1998 to June 2010 were retrospectively analyzed.According to the condition of bile duct stones,laparoscope,choledochoscope and duodenoscope were applied separately or combinately.All patients were divided into A,B and C groups.Patients in group A were treated by laparoseopy,choledochoscopy or duodenoscopy;patients in group B were treated by choledochoscopy+duodenoscopy.duodenoscopy+laparoscopy or laparoscopy+choledochoscopy;patients in group C were treated by laparoscopy+duodenoscopy+choledochoscopy.The efficacies of different treatment approaches were analyzed by comparing the results of imaging examination and follow-up.Results The curative rate and complication rate of the group A were 89.54%(1276/1425)and 6.73%(86/1276),respectively.Of the 149 patients in the group A who were failed in the treatment,83 patients were transferred to the group B,and 66 patients were transferred to the group C.The curative rate and complication rate of the group B(including 83 patients transferred from the group A)were 95.93%(1719/1792)and 4.07%(70/1719),respectively,and 73 patients who were failed in the treatment were transferred to the group C.The curative rate and complication rate of the group C(including 139 patients transferred from the group C)were 99.75%(783/785)and 0.26% (2/783),respectively,and 2 patients who were failed in the treatment received open surgery.Conclusion Combined application of laparoscope and endoscopes could raise the success rate of stone clearance and decrease the postoperative complications.

Objective To establish diagnostic standards of confocal laser endomiroscopy (CLE) in pathological classifications of gastric cancer and evaluate the diagnostic value of CLE in vivo. Methods The confocal images of 36 patients, who were diagnosed as gastric cancer with confirmation of pathology, were collected and analyzed to establish diagnostic standards of CLE for pathological classifications of gastric cancer. The sensitivity, specificity and accuracy of CLE were determined with reference to histopathologic findiugs retrospectively. Results According to changes of glandular architecture and microvessels under CLE, gastric cancers were divided into differentiated and undifferentiated type. The sensitivity, specificity and accuracy of CLE in diagnosing differentiated carcinoma were 85.7%, 92. 3% and 90. 5%, respectively, which were 85.7%, 92. 3% and 90. 5%, respectively, in undifferentiated carcinoma. Conclusion CLE is a new tool to make pathological classifications of gastric cancer during routine endoscopic examination procedure, which correlates well with histopathologic fingdings.

Objective To estimate the value of confocal laser endomicroscopy (CLE)for differential diagnosis of gastric ulcer. Methods Patients diagnosed as gastric ulcer by conventional endoscopic mode of CLE were enrolled. The eonfocal endoscopic mode of CLE was used to observe the borderline and/or the surface of the ulcers. Biopsy specimen was taken for histopathological examination and the result of CLE was compared with histopathological finding. Results A total of 12 769 CLE images were obtained from 150 sites of 43 patients. The sensitivity and specificity of CLE for phlegmonosis process were 83.54% and 85.92%, respectively, with positive predictive value of 86.84% and negative predictive value of 82.43%. Whereas the sensitivity and specificity of CLE for intraepithelial neoplasia were 53.57% and 88.52%, respectively, with positive predictive value of 51.72% and negative predictive value of 89.26%. As for diagnosis of gastric cancer, the sensitivity and specificity of CLE were 88.37% and 93.46%, respectively, with positive predictive value of 84.44 % and negative predictive value of 95.24%. The kappa values for agreement of histopathological findings and CLE for diagnosis of phlegmonosis process, intraepithelial neoplasia and gastric cancer were 0.69,0.42 and 0.81, respectively. Conclusions CLE can provide a real-time histopathological diagnosis of the gastrointestinal tract with high sensitivity, specificity, positive and negative predictive values. Targeted biopsy can be done under CLE and the detective rates of precancerous lesions and tumors will be increased.

Objective To copy the mouse aging model with D-galactose,and to investigate the role of Schisandra total lignin (SCL)in the mouse brain tissue aging and its mechanism.Methods 50 mice were radomly divided into control group,model group (100 mg·kg-1 ·d-1),low dose (35 mg·kg-1 ·d-1)of SCL group (SCL-L), middle dose (70 mg· kg-1 · d-1 )of SCL group (SCL-M)and high dose (140 mg· kg-1 · d-1 )of SCL group (SCL-H)(n=10).D-galactose (100 mg·kg-1 ·d-1 )was injected into the mice hypodermically for 10 weeks to induce aging models in all the groups except control group,and 35,70,and 140 mg· kg-1 · d-1 SCL were administered for 10 weeks in SCL groups.The learning and memory abilities were measured by the Water Maze test.The expression levels of Bax,Bcl-2,ubiquitin (Ub),microtubule-associated protein light chain 3 (LC3)in the brain tissue of the mice in various groups were observed by Western blotting method. The LC3 protein expressions in mouse brain cortex and hippocampus were observed by immunohistochemistry.Results In learning and memory test,compared with control group,the swimming time of the mice in model group was increased (P<0.05),and the number of errors was increased (P<0.05);compared with model group,the swimming time in SCL-L,SCL-M and SCL-H groups was decreased (P<0.05)and the number of errors was also decreased (P<0.05). Compared with control group,the expression level of Bax was increased (P<0.05),the expression level of Bcl-2 was decreased (P<0.05),the expression levels of Ub and LC3-Ⅱ/LC3-Ⅰ proteins were increased (P<0.05)in model group;compared with model group,the expression level of Bax was decreased (P<0.05),the expression level of Bcl-2 was incerased (P<0.05),and the expression levels of Ub and LC3-Ⅱ/LC3-Ⅰ proteins were decreased (P<0.05)in SCL-L,SCL-M and SCL-H groups.In control group,the neuronal morphology was normal,and none of brown granules were visible in the cytoplasm of mouse brain cortex and hippocampus and the expression of LC3 protein was negative.In model group,the neurons were degeneration,and the number of LC3 protein positive cells in the cerebral cortex and hippocamptal tissue was increased (P<0.05).In SCL-L,SCL-M and SCL-H groups,the number of degenerative neurons was decreased,and the number of LC3 protein positive cells was decreased (P<0.05).Conclusion SCL can inhibit the D-galactose-induced brain tissue aging in the mice, and the mechanism is related to regulating autophagy and inhibiting apoptosis.

ObjectiveTo investigate the treatment of Mirizzi syndrome (MS) by ERCP、laparoscopy and choledochoscopy. MethodsIn this study 12 cases were confirmed intraoperatively as with MS from July 2005 to June 2009. Patients were treated by ERCP,laparoscopy and choledochoscopy according to the Csendes Classification. ResultThere were 7 MS patients complicating common bile duct stones among all 12 MS cases.There were 8 cases of Type Ⅰ Csendes MS,3 cases of Type Ⅱ and 1case of Type Ⅲ.11cases were treated by the ERCP、laparoscopy and choledochoscope.4 cases was treated by primary closure of common bile duct in laparoscopy,all the patients were cured.The case of type Ⅲ with T tube placed for stone caused bile duct injury had no stricture of the common bile duct as demonstrated by postoperative follow-up cholangiography.ConclusionsERCP,laparoscopy and choledochoscopy are effective for the treatment of Mirizzi syndrome.

Objective:To evaluate the effectiveness of integrated pediatric training course of extracorporeal membrane oxygenation(ECMO)based on simulation teaching.Methods:The instructors of ECMO team of Children′s Hospital of Fudan University developed the teaching curriculum.Team members as a unit were recruited to study simulation-based courses, whose theoretical knowledge, skills and teamwork were assessed, and feedback from the trainees were gathered.Results:Since May 2018, 22 teams of pediatric specialized hospitals or general hospitals have taken part in our ECMO simulation, with totally 132 participants, including 45 ICU doctors(34.1%), 60 ICU nurses(45.5%), 23 thoracic surgeons(17.4%) and four anesthesiologists(3.0%). Twelve of them(9.1%)had a little ECMO experience.After training, the trainees had a high evaluation on each part of the course, with average score more than four, of which the theoretical score was lowest.Their self-evaluation on the theory, skills, teamwork and confidence in the implementation of ECMO had been significantly improved.Most(80.3%)of the trainees were confident to carry out ECMO in their local hospitals.The trainees′ baseline score of theoretical knowledge was 55.2±7.6, and increased to 67.1±7.3 after training( P<0.001). The average teamwork score of the 22 teams was 70.2±8.2, and the qualified rate was up to 86.4%.The most prominent skill problems in operation assessment were no albumin and blood priming(90.9%), non-skill problems were extremely anxious during the overall assessment(84.8%)of the participants, poor team work(74.2%), poor leadership(68.2%). For the feedback of the whole course, 97.0% and 94.7% of the trainees thought that integrated training and simulation teaching were the highlights of the course.At present, 13(59.1%)units have successfully developed ECMO technology after the course, and the number of children treated has been up to 83, with a survival rate of discharge of 54.2%.However, it was found that the skill level of some participants decreased about 2 weeks after the course. Conclusion:As a new attempt of ECMO teaching, this curriculum has achieved certain efficiency in both teaching and clinical practice.At the same time, our study also plays a role in promoting the development of ECMO in pediatrics.

Objective To assess the value of blood oxygen level-dependent magnetic resonance imaging (BOLD-MRI)in diagnosis and prediction of early acute renal transplant rejection.Methods BOLD-MRI was performed in a cohort of 103 patients undergoing cadaver renal transplantation between Dec 2005 and March 2007.Among them,82 recipients had nomlal renal function,21 had biopsy-proved acute rejection.R2* (1/s)measurements were obtained in the medulla and cortex of grafted kidneys. Results R2* values of the medulla were significantly lower in the acute rejection group[R2*=(14.02±2.68)/s]than that in the normally functioning transplants group [R2*=(16.66+2.82)/s],the difference between these two groups was significant (P＜0.01);ROC curve analyses suggested that medullary MR2* values could accurately identify acute rejection in the early post-transplantation period.In the normal functioning transplant group,those with lower medullary R2* values (MR2*＜14.9/s,n=23) had higher acute rejection rates than those with higher medullary R2* values (MR2*＞14.9/s,n=59) in the first 6 months following transplantation,but the difference between these two groups was not significant (17.39% vs 8.47%,P=0.259). Conclusions Mean R2* values in the medullary regions of grafted kidneys with BOLD-MPd may be a non-invasive diadynamic criteria with good sensitivity and specificity,and may be a valuable predictor of early acute renal transplant rejection.