Objective To establish better reaction factors for rep-PCR and to investigate if the extended spectrum ?-lactamase(ESBL)+ Escherichia coli isolated from different patients in the respiratory ward have the same origin by using rep-PCR.Methods The ESBL confirmation was taken by double disc confirmatory test.The susceptibility testing was performed with K-B test.By applying the widespread enterobacter repetitive intergenic consensus as a primer,the stains were typed by rep-PCR following electrophoresis in agarose gel.Results The analysis of the PCR productions indicated that it would create useful DNA ban-prints and all these 22 ESBL(+)Escherichia coli strains were of three origins.Conclusion The method of rep-PCR is practical for epidemiological research in nosocomial infection.

Objective To investigate the clinical value of fecal calprotectin as a marker for diagnosis of ulcerative colitis(UC).Methods ELISA assay was used to measure the concentrations of calprotectin in feces of 30 UC patients and 20 healthy controls.The disease activity of UC was determined by Truelove Witts histological criteria.Results The level of calprotectin in feces was significantly higher in feces with active UC(578?g/g) than with inactive UC(8.4?g/g,P0.05),but significant difference existed between each active grade and inactive phase of UC.The results revealed siginificant correlation between the endoscopic grades of disease activity and fecal calprotectin concentration(r=0.85,P

Objective To construct a recombinant lentiviral vector for HCV core-Ant and then study its effect on the transdifferentiation of hepatic stem cells. Methods The HCV core-Ant was obtained by PCR of two primers template with each other and T-vector cloning method,and then subcloned to pLenti6/V5-D-TOPO. The restriction endonuclease and T4 DNA ligase were used to construct the vector. pLenti6/V5-D-HCV core-Ant and the ViraPowerTM PackagingMix (containing three packaging plasmids pLP1,pLP2 and pLP/VSVG) were cotransfected into 293ET cells to produce replication in competent lentivirus after transfection. The viral supernatant on 293T cells was collected. The expression of the lentiviral vector containing HCV core-Ant in Hela cells was measured by immunohistochemistry. Then we constructed the lentiviral vector containing green flurosecent protein by the similar method,and the titers were determined. Results HCV core-Ant was identified and analyzed by restriction enzyme digestion and sequencing,respectively. The expression of the recombinant lentiviral vector plasmid containing HCV core-Ant in Hela cells was confirmed by immunohistochemistry. The 3T3 cells transfected the lentiviral vector containing green flurosecent protein were found to show strong expression of GFP,which confirmed that the four-plasmid system of the lentiviral vector was successfully constructed. Conclusion The recombinant lentiviral vector expressing HCV core-Ant was successfully constructed by molecular cloning and recombination techniques in vitro,which will be beneficial to guiding further study on gene therapy of cancer.

Objective Try to know the prevalence rate of ESBL-s produced by Eseherichia coil in our hospital from 2003 to 2007.Methods Antibiotic susceptibility were determined by K-B disc diffusion.Double-disk synergy tests performed on Mueller-Hinton agar plates was used for ESBL confirmation.Results There was 23.8% of our isolates can produce ESBLs in 2003,25.4% in 2004,31.3% in 2005,36.6% in 2006 and 44.9% in 2007,respectively.The situation of antibiotic resistance is more severe.Conclusion The antibiotic resistance should be well surveyed and the ESBLs must be routinely assayed.

Objective To assess the differences in the specificity and the sensitivity in detecting specific IgM by indirect ELISA and capturing-ELISA methods,and to analyze the possible causes of those differences.Methods The HBc-IgM and EHF-IgM level in serum samples from activity Hepatitis B patients and hemorrhagic fever patients with renal syndrome were determined by both indirect-ELISA and capturing-ELISA methods,and the differences in the positive rate,positive threshold value and the specificity between those two methods were compared. Results Specific murine IgM diluted in PBS were detected by indirect-ELISA and capturing-ELISA,and the specificity and sensitivity of those two methods were similar.Results showed that sensitivity of indirect-ELISA was lower than capturing-ELISA in detecting specific IgM in serum from patients with activity Hepatitis B and hemorrhagic fever patients with renal syndrome.The IgM level in hepatitis B and hemorrhagic group treated with thioglycol became negative detected by those two methods,suggesting that both of them have the anti-IgM epitope-specificity.Cross reaction results demonstrated that the reagent detecting IgM from the two methods had the specificity to the specific antigen. Conclusion It is recommended to detect IgM level by capturing-ELISA method for the early diagnosis of acute infectious disease,pathological changes,immune reaction and prognoses of chronic continuous infectious disease.Applying the indirect-ELISA method to detect IgM level to diagnose acute infectious disease is to discuss in the future.

Objective To evaluate the ability of further discriminating diagnosis of the headchest lead electrocardiogram (HCECG) in elderly patients with abnormal Q waves in routine lead electrocardiogram (RLECG) in inferior lead. Methods The 55 male patients, aged 65-88 years,with abnormal Q waves in both lead Ⅲ and aVF were selected and divided into two groups: myocardial infarction (MI) group and non-MI group, according to if the patient had a history of acute MI. All the patients accepted examination of coronary computed tomographic angiography (CTA) and ultrasound cardiogram, those with Wolff-Parkinson-White syndrome and myocardial hypertrophy were excluded.The 30 individuals of control group had no abnormal Q waves in lead Ⅱ , Ⅲ and aVF. HCECG and RLECG were recorded simultaneously in respective groups and occurrence rate of Q waves in correlative leads Ⅱ, Ⅲ, aVF and HL3, H0, HR3 were matched and compared, sensitivity and specificity were compared in respective leads. Results There were serious stenosis in 22 patients (100%) in MI group, and there were mild stenosis in 10 (30. 3%) and moderate stenosis in 23 patients (69.7%) in non-MI group. There was no significant difference between HCECG and RLECG in occurrence rate of Q waves in MI group (P＞ 0. 05 ). Non-MI group left anterior axillary line, Ⅱ ,Ⅲ, right anterior axillary line, near the anterior midline, aVF without Q wave and exclusion of old MI diagnostic specificity were 100%, 97.0%(32/33), 97.0% (32/33), 15.2% (5/33), 100%, 39.4%( 13/33)respectively. Conclusions Pseudo-changes are rarely found in HCECG and there is a higher degree of conformity in HCECG with coronary lesions, therefore HCECG may be used to discriminate whether the inferior abnormal Q waves occurred in RLECG are truly abnormal or not.

Objective To investigate the related factors of percutaneous balloon compression (PBC) for the treatment of trigeminal neuralgia complicating cerebrovascular disease in order to reduce the incidence of surgical complications. Methods The clinical data in patients with trigeminal neuralgia complicating cerebrovascular disease treated with PBC in the People's Hospital of Liaoning Province from December 2000 to December 2010 were analyzed retrospectively. The reasons for its occurrence were investigated. Results Of the 6 759 patients with trigeminal neuralgia who received PBC treatment, 10 (0. 15％ ) developed cerebrovascular disease,and 2 of them had arteriovenous fistula; 5 patients developed intracranial hemorrhage: 2 acute subdural hemorrhage, 2 intraventricular hemorrhage with subarachnoid hemorrhage, and 1 small amount of bleeding in brain stem; 1 patient developed internal carotid hemorrhage during the intraoperative puncture and the operation was stopped temporarily, and he was reoperated successfully after 2 weeks; 2 patients had cerebral infarction, they were all in the risk population of cerebrovascular disease, and 3 weeks after treatment, their disease conditions were improved. One of them left severe paralysis of limbs at discharge. Condusions The treatment of trigeminal neuralgia with PBC is safe and the incidence of cerebrovascular complications is lower. The surgical operations and blood pressure changes are the important causes of cerebrovascular complications.

Objective To analyze composition principles of prescriptions for the treatment of diarrhoea caused by deficiency of spleen. Methods The prescriptions for diarrhoea caused by deficiency of spleen in the Prescriptions of Traditional Chinese Medicine Dictionary were collected, sorted and entered into the TCM Inheritance Support System (V1.3) to analyze the composition principles through the methods of entropy clustering and apriori. Results Based on the analysis of 1185 prescriptions and 815 medications, there were 33 medications with more than 50 frequencies. Composition principles were obtained through apriori method:50 herbal pairs were used with more than 50 frequencies, and 29 core combinations with more than 40 frequencies. Association principle rule were used for the analysis of those medications in the prescriptions (support≥20%, confidence≥0.9). Principles were obtained through entropy clustering:there were 19 core combinations which composed the new prescriptions, and 19 new prescriptions were found through hierarchical clustering method. There were 17 prescriptions matching with Sijunzi decoction (semblance=0.5). Conclusion Diarrhoea caused by deficiency of spleen should be treated with strengthening spleen and benefiting qi primarily, assisting with warming yang, excreting dampness and checking diarrhoea.

Objective To develop a quantitative immunohistochemistry assay for duck hepatitis B virus core antigen (DHB-cAg)in duck liver tissue.Methods By comparison with no repair antigen and repair antigen with high pressure,microwave and trypsin,the best solution of antigen retrieval was determined.By optimizing the parameter of image acquisition and de-ducting blank area,mean density of yellow areas was calculated using Image-Pro Plus 6.0 software.Using the assay devel-oped to determine the level of DHBcAg in liver tissue from duck infected by DHBV,anti-DHBV activity of DHBcMAb-TAT PTD conj ugate was examined.Results SABC method with no repair antigen was selected,which was better than other methods.DHBcAg expression in duck liver tissue could be objectively and accurately quantified by setting Image-Pro Plus 6.0 software parameters and calculating mean density of yellow areas.By comparison with the differences between mean densityat baseline of treatment and end of treatment,it was showed that DHBcMAb-TATPTD conjugate treatment dose-de-pendently reduced the levels of DHBcAg in liver tissue,which show that the assay developed could effectively evaluate the anti-DHBV activity of agent.Conclusion The immunohistochemistry assay developed in this study can objectively and accu-rately evaluate the level of DHBcAg in duck liver tissue.

Objective To explore the timing of surgical operation and surgical incision for the treatment of head of electrical injury and scarred alopecia with soft tissue expander .Methods Twenty-three patients with high tension electrical injury and scarred alopecia were collected ,the soft tissue expander was used to repair the wound and scar area.Selected the expander to embed the incision:patients with scarred alopecia were selected to be near the hair area in the scar;high tension electrical injury in patients with incision location selection based on the time of injury and wound repair requirements vary;within 5 days after injury ,surgery incision can be selected from the wound about 2-3cm;more than 5 days after injury,the patients chose to stay away from the wound incision ,incision about 4-5cm. Results Of 23 patients,postoperative scalp expansion flap good blood supply ,a period to solve the high tension electrical injury caused by skull necrosis and scarred alopecia problems .The patients according to the wound debride-ment and repair requirements using different surgical incision for dilator implantation ,wound and flap infection ,dilated flap poor blood supply ,necrosis,dilator exposed and other issues did not occur after surgery .Conclusion Flexible use of expander embedded surgical incision can be more effectively adapted to the wound repair requirements ,timely reduction of implantation expander surgery when the difficulty of stripping ,shorten the operation time,the maximum possible protection of healthy scalp tissue ,and eventually get a satisfactory clinical cosmetic repair effect .