Objective To develop and verify a double antibody sandwich ELISA detection method for the determination of ovalbumin(OVA),in order to determine the OVA content in influenza vaccines.Methods The rabbit anti-OVA polyclonal antibody was used as coating antibody and HRP labeled rabbit anti-OVA polyclonal antibody as detection antibody to develop a double antibody sandwich ELISA for OVA.The antibody concentration(2,1,0.5 and 0.25 pg/mL) for coating,enzyme-labeled antibody concentration(0.5,0.25 and 0.125 μg/mL),and kinds of blocking reagent(blocking with1% BSA,blocking with 2% BSA,blocking with 1% BSA and 1% sucrose,and blocking with 2% BSA and 2% sucrose,using nonblocking as control) were optimized,and the Cut-off value was determined as the judgment standard.The developed method was verified for the linear range and detection limit,specificity,repeatability and accuracy.Results The optimum detection conditions were as follows:the concentration of coating antibody was 1 μg/mL,the concentration of enzyme-labeled antibody was 0.25 μg/mL;The blocking reagent was 2% BSA and 2% sucrose;The Cut-off value was 0.051 66.The linear range of the method was 5～0.313 ng/mL,and the detection limit was 0.078 ng/mL;It did not react with influenza virus,bovine serum albumin(BSA) and Vero cell supernatant;The intraplate and interplate coefficient of variation(CV) were between 2.562%～13.887% and 4.000%～16.497% respectively;The coincidence rate between the results of this method and the Germany SERAMUN OVA quantitative test kit ranged from 93.79% to 107.05%.Conclusion The developed OVA double antibody sandwich ELISA has good specificity,repeatability,linear range and accuracy with convenience and lower cost,which might be used for the quantitative detection of OVA.

Electromagnetic Fields ; Environmental Exposure ; Humans ; Radiation Dosage ; Threshold Limit Values

Objective To explore the status and influencing factor of psychological insulin resis tance (PIR) of type 2 diabetes.Methods 137 in-patients with type 2 diabetes were investigated by selfdesigned questionnaire to get general data and Insulin Perceptions Questionnaire to get PIR data.Multiple linear regression were used to find influencing factor.Results 134 participates had PIR among the 137 in-patients.The older of age,using insulin for longer time and receiving insulin treatment positively had the higher scores of main questionnaire,then had less serious psychological insulin resistance.The duration of illness course,complications,work and using insulin for the first time or not had no relationship with their PIR.Conclusions The findings indicated that the type 2 diabetes using insulin have psychological in sulin resistance generally.Age,the time using insulin,accepting or receiving insulin positively are the independent factors of psychological insulin resistance

Objective To explore the X-ray and MRI manifestations of hip joint lesion in ankylosing spondylitis patients and give useful reference for diagnosis,assess of disease activity and therapeutic response and prognosis.Methods The X-ray manifestation and MRI manifestation of 64 cases of patients with ankylosing spondylitis were analyzed.Results In X-ray analyzed acetabulum joint changes 42,hip joint hyperostosis 47,hip joint interval narrow 13,hip joint osteoporosis 11,MRI analyzed synovial inflammatory change 64,subcartilaginous bone marrow edema 48,joint cartilage lesion and joint surface saccular change 42,tendon and ligament of enthesopathy 24,joint space change 13.Conclusion In patients with ankylosing spondylitis,X-ray manifestation of hip-joint lesion commonly included leacetabulum joint changes,hip joint hyperostosis and hip joint space narrowing while MRI manifestation included inflammation of synovium,edema of subcartilaginous bone marrow,and tendon of enthesopathy which suggested high disease activity.To suspicious hip-joint lesion in patients with ankylosing spondylitis,magnetic resonance imaging should be taken for judgment of disease activity and determine of treatment plan.

Pancreatic cancer is one of the worst digestive malignancies characterized by non-specific symptoms, rapid progression and high mortality. Despite great efforts that have been made in basic and clinical research, the prognosis remains poor with an overall 5-year survival rate of less than 5%. Complete surgical resection is the only curative treatment option, but the curative resectabillty is still about 15% in China. Pancreatic surgery is considered one of the most techni-cally demanding and challenging procedures. The development of pancreatic surgery in China depends not only on the progress in surgical techniques, but also the specialization on pancreatic surgery. There has been advancement in combined therapy with a modern interdisciplinary approach including chemotherapy,radiotherapy, biotherapy and thermotherapy. The research pro-gress of gene therapy on pancreatic cancer showed us some hopes for the future. The outcome of pancreatic cancer treated by Chinese medicine with herbal drugs is encouraging, but still needs the support of more solid evidences from randomized con-trolled trials. It is suggested that the combined therapy should play an important role in the treatment of pancreatic cancer, and the fallow-up should be paid more attention.

Objective To evaluate the relationship between functional oipA gene of Helicobacterpylori and digestive disease. Methods Biopsy gastric mucosa were obtained from 360 patients who would get gastroscopy.Hp were isolated from urease positive samples and partly urease negative samples,and improved by microscope,urease experiment and 16SrRNA PCR. OipA and cagA of the isolates were obtained by PCR and the statas of oipA signal region were analysised after sequencing. Analyze the relevance between functional oipA gene of Helicobacter pylori and digestive disease.Results 106 isolated Hp were obtained,in which 72 strains with cagA gene positive,87 with oipA signal region gene positive (80 the statas of signal region was on and 1 was off;6 could not be decided).The frequency of functional oipA were both significantly higher than cagA in ulcer and atrophic gastritis.Conclusion The relationship between functional oipA gene and digestive disease was more closer than that of cagA.

Objective To establish an indirect ELISA method with Helicobacter pylori (HP)recombinated antigene OipA6 as coating antigen after prokaryotic expression and purification.Methods The ELISA plate was coated with Hp OipA6 recom-binant protein as antigen.Serum specimens and enzyme labeled anti human IgG were added.The reaction conditions was op-timized.The results of indirect ELISA were compared with the detection of oipA gene.Results When Hp OipA6 recombi-nant protein was 1.0 mg/ml,the condition of coating was 2~8℃ one night,the dilution of serum specimens was 1∶100 and enzyme labeled anti human IgG was 1∶4 000,the incubation time of enzyme labeled anti human IgG was 40 min,the chro-mogenic time was 15 min,the result of ELISA was the best.The sensitivity and specificity of the indirect ELISA were both higher than the detection of oipA gene.Conclusion It suggested that the indirect ELISA,in which Hp OipA6 recombinant protein were coated,may be an appropriate assisted method in detection of high virulent Hp infection.

Objective To explore the effectiveness of insulin dialogue toolkit on psychological insulin resistance of type 2 diabetes. Methods A total of 134 type 2 diabetes with psychological insulin resistance were selected by the inclusion and exclusion criteria . The participates were randomly assigned to control group and intervention group by computer with 67 cases each. The control group was given conventional health education, the intervention group was given insulin dialogue toolkit instead of individual health education based on conventional health education. The intervention was lasted 2 weeks.The total score and each dimension score of my opinion of insulin and the blood glucose before and after intervention were observed. Results Before intervention, the total score in my opinion of insulin in intervention group and control group was (66.40±8.51), (66.91±7.27) scores, and there was no significant difference between two groups, P>0.05.After intervention, the total score in my opinion of insulin in intervention group and control group was(76.93±5.31), (71.09±7.77) scores, and there was significant difference between two groups, Z=-5.585, P<0.01. There was significant difference in each dimension score, P<0.05 or 0.01. After intervention, the fasting blood glucose in intervention group and control group was (7.35±1.76), (7.54±1.70) mmol/L,the postprandial blood glucose 2 h in intervention group and control group was (9.47±2.26), (8.79±2.04 ) mmol/L, and there were no significant differences, P>0.05. Conclusions The insulin dialogue toolkit has an impact on psychological insulin resistance of type 2 diabetes. Insulin dialogue toolkit is a way to improve the diabetes′attitude of insulin, but the effect on blood sugar are not clarity.

Objective:To explore the effect of the labor abdominal pressure training device on the second stage of labor and the outcome of labor in primiparas undergoing epidural analgesia.Methods:The healthy primiparas who had patient controlled epidural analgesia were randomly divided into the observation group (126 cases) and the control group (122 cases). The control group used conventional oral instruction to force after entering the second stage of labor. The women in the observation group used the labor abdominal pressure training device to guide the force. The time required to learn to exert force, the second stage of labor and the outcome of labor were observed.Results:The parturient women learned the correct time to exert force, the time of the second stage of labor, the self-efficacy score of the second stage of labor, and the post-natal fatigue score were (13.56 ± 5.32) min, (76.32 ± 10.58) min, (121.38 ± 17.52)points, (8.56 ± 0.72) points in the observation group, and (25.47 ± 7.65) min, (105.21 ± 18.41) min, (101.42 ± 14.32) points, (13.31 ± 0.46) points in the control group, the differences between the two groups were statistically significant ( t values were -10.326-11.002, all P<0.05). The maternal spontaneous delivery rate, fetal assisted delivery rate, forceps assisted delivery rate, perineal lateral cut rate, perineal median cut rate, perineal tear>Ⅰ°rate were 94.44% (119/126), 4.76% (6/126), 0.79% (1/126), 9.52% (12/126), 11.11% (14/126), 6.35% (8/126) in the observation group, and 85.25% (104/122), 12.30% (15/122), 2.46% (3/122), 29.51% (36/122), 4.10% (5/122), 9.84% (12/122) in the control group,the differences between the two groups were statistically significant ( χ2 values were 4.582-9.437, P<0.05 or 0.01). Conclusions:The use of an abdominal pressure training device for labor in patient controlled epidural analgesia for pregnant women can shorten the second stage of labor and study the time of exertion, improve the self-efficacy of the second stage of labor, reduce the degree of perineal injury and fatigue, reduce the vaginal birth rate, and help protect Mother and child are safe.

The expressions of survivin,PcNA and Bcl xl were detected in 68 patients with pramary hepatic careinoma(HCC) and adjacent tissues and 12 cases of normal hepatic tissues by immunohistochemical (IHC) EnVision and Tunel method.The relationship between survivin gene expression and apoptosis index(AI),proliferation index(PI) and apoptosis inhibition factors were studied.The positive rate of survivin and Bcl xl expression were 58.8%(40/86) and 48.5%(33/68) in HCC,5.88%(4/68) and 36.8%(25/68) in adjacent tissues,0 (0/12) and 25%(3/12) in normal hepatic tissues.The expression of Survivin and Bcl xl are mainly in cell membrane and cytoplasm.Survivin expression is related to metastasis of HCC ( P