Objective:To construct an immune inhibitor of new TNF-?,C-terminal sequence of new rhTNF-? was replaced with sequence of T-help cell of hen egg-white lysozyme(HEL).Methods:The rhTNF-? mutant was cloned 、expressed and purified.Results:The DNA sequencing analysis showed that the C-terminal sequence of new rhTNF-? mutant was correct.The mutant was inserted into pBV220 expression vector .After the recombinant bacteria was incubated at 42℃ for 4 h,a new band of the protein with relative molecular weight of 1.7 kD was shown on the gel.The band amounted to 30 % of total bacteria protein.Western blot showed that the mutant protein could associated with anti-TNF-? antibody.After the protein was purified by through a column of Q-Sepharose Fast Flow, the purity of the protein was above 90%.The biological activity of the protein was measured with L929 cells.The result showed that biological activity of the protein was totally lost.Conclusion:The experimental evidence demonstrated that the construction of new rhTNF-? mutant was successful.The mutant not only can be associated with anti-TNF-? antibody, but also lose the biological activity of original TNF-?. [

0.05). PetCO 2 was a reliable index reflecting PaCO 2. There was a significant difference between PetCO 2 and PaCO 2 in the Group Ⅲ (P

Objective: To investigate the influence of high-fat diet on glucose metabolism and fatty acid composition of serum phospholipid in OLETF rats, a model of spontaneous type 2 diabetes mellitus. Method: Twenty male OLETF rats of 14 w age were divided into 2 groups randomly. The dietary fat in two groups contained identical percentages of saturated fatty acids, monounsaturated fatty acids and polyunsaturated fatty acids. Oral glucose tolerance test and measurement of insulin level were performed after 10 w treatment. The fatty acid composition of serum phospholipid in rats was observed as well. Results:The food intakes were comparable in 2 groups. The body weight gain in high-fat group was higher than standard group during the experiment. The result of oral glucose tolerance test and measurement of insulin level showed that the glucose metabolism in two groups had no significant difference. Only the percentage of total n-6 polyunsaturated fatty acids, 18:3(n-3) of serum phospholipid were comparable, percentages of other fatty acids were significantly different in two groups. Conclusion:With the comparable fatty acid composition, higher dietary fat lead to more body weight gain, but had no significant influence on glucose metabolism in OLETF rats. The amount of dietary fat had significant effect on fatty acids composition of serum phospholipid in OLETF rats.

Objective To study factors influencing injury severity of expressway traffic acci- dents. Methods Data of expressway traffic accidents involving casualties in areas around Hangzhou during 2005-2006 were collected and analyzed by non-conditional simple and multiple logistic regression analysis. Results (1) During 2005-2006, there were 220 traffic crashes involving 327 eases, of which there were 64 deaths (19.6%), 121 severe injuries (37.0%, ISS≥16 points) and 142 slight and mod- erate injuries (43.4%, 1 point≤ ISS≤15 points). (2) Non-conditional simple logistic regression analy- sis showed that risk factors significantly influencing the injury outcomes of traffic accidents included gen- der, weekday, weather, type of collision, vehicle type, driver category, purpose of vehicle use, purpose of travel and crash responsibility. (3) Non-conditional multiple logistic regression analysis manifested that influential factors were gender, type of collision, driver category and crash respensibihty. Conclu- sion Analysis of expressway traffic accidents data helps identify factors influencing injury severity of ex- pressway traffic accidents may provide suggestions for prevention or decrease of expressway traffic acci- dents.

Objective To investigate the relationship between the X-ray,CT findings and its pathological basis of nonossifying fibroma(NOF).Methods X-ray and CT features of NOF confirmed by surgical pathology in 17 cases,and correlative study of imaging andpathology was done.Results 17 lesions were showed as eccentric and expanded destruction located in the cortex,and the remains of bone could be found inside the tumor,the lesions projecting into marrow cavity with a thin bone between the tumor and marrow.The tumors were grey-yellow or brown tissue around by the thin bone found in operation.NOF was consisted of a lot of shuttle cell and collagen fiber under microscope,some of macrophagocyte with multiple nucleuses and foam cell dropped disseminating in them,and there was not osteogenesis in the tumor.Conclusion The pathological changes of NOF can be reflected by X-ray and CT.

Objectives To investigate the correlation between HIF-1α expression level and clinical pathological characteristics of ovarian carcinoma in the elderly.Methods The expression levels of HIF-1α in normal ovary tissue,benign ovarian tumor tissue,borderline ovarian tumor tissue and ovarian cancer tissue were detected in the elderly by immunohistochemical method.And the correlations between HIF-1α expression level and clinical pathological characteristics of ovarian carcinoma were analyzed in the elderly.Results The HIF-1α expression was almost nothing in normal ovarian and benign ovarian tumor tissues,low in the borderline ovarian tissues,and high in the ovarian cancer tissue(Z=11.324,P =0.002).The IOD value of tissues was higher in borderline ovarian tumor tissue[(2.58 ± 0.65) × 105] and ovarian cancer tissue [(5.95 ± 1.02) × 105] than in normal ovary tissue[(1.90±0.35) × 105]and benign ovarian tumor tissue[1.95±0.41) × 105] (F=15.711,P=0.001).The expression of HIF-1α in the tissues was lower in ovarian cancer with stage Ⅰ-Ⅱ,high or moderate differentiation and non-metastatic lymph node than in ovarian cancer with stage Ⅲ-Ⅳ,low differentiation and metastatic lymph node (x2 =4.714,6.121,6.364;P =0.030,0.013,0.012).Conclusions The expression of HIF-1α is up-regulated in ovarian cancer in the elderly compared with normal ovarian tissues,which may be related to occurrence,development and clinical prognosis of ovarian cancer.

Objective To compare the effect between problem-based learning and lecture-based learning for clinical teaching in the department of cardiovascular medicine. Methods Totally 110 five-year-program cardiovascular interns from June 2011 and June 2012 were selected. They were randomly divided into the PBL group(n=55) and LBL group(n=55). PBL and LBL teach-ing methods were applied in the two groups respectively. Teaching effects were evaluated by exam and questionnaire investigation. SPSS 15.0 was used to do data processing; t test was used to compare the average score of two groups;chi-square test was used to process the results of the questionnaire. P<0.05 signifies sta-tistically significant differences. Results There were statistical differences in examinational average score between PBL group and LBL group ((87.89 ±5.39) vs. (82.63 ±5.26), P<0.05). PBL group had significantly higher satisfaction rate in motivating study interests , deepening understanding of theoretical knowledge, cultivating self-learning ability, training verbal expression and developing clin-ical thinking, etc(P<0.05). Conclusions PBL teaching method demonstrates advantages in teaching of cardiovascular medicine and enhances the teaching effect. But the PBL teaching method should be improved in basic knowledge teaching, cultivation of teachers' ability and case selection.

Objective To explore the clinical manifestations and recovery from neuroimaging abnormalities of acute disseminated encephalomyelitis (ADEM),in order to improve the understanding of ADEM.Methods A total of 47 children with ADEM in Department of Neurology,Children′s Hospital of Soochow University were followed up for 1 year.The clinical manifestations,auxiliary examination (especially neuroimaging),treatment and prognosis were analyzed.The recovery conditions from clinical and neuroimaging abnormalities were summarized.Results Among 47 cases,37 cases (78.72%) had prodromic infection history,and 5 cases (10.64%) had the history of vaccination.The clinical manifestations were varied,including 36 cases (76.60%) with somatic discomfort,30 cases (63.83%) with fever,30 cases (63.83%) with disturbance of consciousness,28 cases (59.57%) with epileptic seizure,24 cases (51.06%) with headache and dizziness,22 cases (46.81%) with nausea and vomiting,15 cases (31.91%) with dyskinesia,15 cases (31.91%) with cranial nerve injury,9 cases (19.14%) with incoordination,and 8 cases (17.02%) with mental and behavior disorders.Abnormal lesions presented multiple,asymmetric patchy and large patchy signal image showed by brain magnetic resonance imaging (MRI),and 7 cases (14.89%) involved the spinal cord.All patients received the treatment of Methylprednisolone [15-20 mg/(kg·d)] combined with intravenous gamma globulin (total 2 g/kg,3 to 5 days).The dose of Methylprednisolone gradually decreased after it had been used for 3 days.In the end,it was replaced by an oral administration of Prednisone [1.0-2.0 mg/(kg·d)],and the total treatment period was for 2-3 months.The follow-ups of 47 children with ADEM showed that the MRI lesions of all the children were reduced and the MRI lesions in 4 (8.51%) of them disappeared completely after 0.5 month of follow-up;the lesions disappeared completely in 13 cases (27.66%) after 1 month of follow-up;the lesions disappeared completely in 23 cases (48.94%) after 3 months of follow-up;the lesions disappeared completely in 32 cases (68.09%) after 6 months of follow-up;the lesions disappeared completely in 43 cases (91.49%) after 1 year of follow-up;and finally there were 4 cases (8.51%) with residual lesions.Conclusion The clinical manifestations of children with ADEM are varied.Most children with ADEM have a favorable prognosis.Imaging lesions of 68.09% of the cases completely disappeared after the onset of 6 months.Imaging lesions in 91.49% of the cases completely disa-ppeared at the onset of 1 year.Only a small fraction of patients present permanent lesions.

OBJECTIVE:To establish an HPLC-CSP (chiral stationary phase) for the determination of plasma levels of enantiomers of fluoxetine. METHODS: Enantiomers of fluoxetine were separated on a Chirobiotic V column. The mobile phase consisted of methanol-acetic acid-triethylamine (100∶0.04∶0.04) with a flow rate of 1 mL?min-1. The detection wavelength was set at 226 nm.Clozapine was used as the internal standard. RESULTS: Under this chromatographic condition, both R-norfluoxetine and S-norfluoxetine had no interference on the enantiomers of fluoxetine and the internal standard. The calibration curve was linear in the range of 8.1～432 ng?mL-1(r=0.999 8) for R-enantiomer and in the range of 10.8～432 ng?mL-1 (r=0.999 2) for S-enantiomer. The relative recovery of R-fluoxetine and S-fluoxetine in plasma were 97.2%～101.9% and 98.7%～102.2%, respectively. RSD of intraday and interday ranged from 2.6% to 5.3% and from 7.7% to 8.9% for R-fluoxetine, and from 2.1% to 6.7% and from 8.0% to 10.6% for S-fuoxetine, respectively. CONCLUSION: The method is simple, reliable and sensitive, and applicable for the determination and pharmacokinetic study of the enantiomers of fluoxetine in plasma.

Objective To reform experimental course teaching methods of clinical bio-chemistry and elevate students interesting and ability. Methods The reformed teaching methods was taken in clinical laboratory techniques and start case-guide comprehensive experiment of clinical biochemistry. Results The innovation of teaching methods of clinical biochemistry obtained satisfactory achievement,improved students thinking skills and overall quality. Conclusion It accorded with the trend of teaching innovation and was advantageous to increasing the comprehensive predisposition.