Objection:To construct and characterize TNFR1-GFP fusion protein expression plasmid(pEGFP-TNFR1) and provide a direct and simplified method for assessment of the effect of TNFR1(siRNA) on the TNFR1 gene expression. Methods:Total RNA was extracted from mouse liver.TNFR1 cDNA was amplified by RT-PCR technique and cloned into pMD18-T vector and the sequence was ensured by sequenciog assay.TNFR1 gene was released from pMD-TNFR1 and subcloned into pEGFP-N2 upstream of GFP gene.pEGFP-TNFR1 was analyzed by restrictive enzymatic digestion to ensure the orientation.The plasmid was then transfected into CHO cells,then the fusion protein expression was detected by immunohistochemistry and fluorescent microscope.Rusults:A 1360 bp gene fragment was obtained and coloned into pMD18-T vector,and the sequence was correct.TNFR1 gene was subcloned into pEGFP-N2 vector,and then restriction endonucleases assays showed the correct orientation.24-48 hours after transfection in CHO cells,the expression of TNFR1-GFP fusion protein can be detected by immunohistochemistry and fluorescent microscope.Conclusion:pEGFP-TNFR1 has been constructed successfully.The TNFR1-GFP fusion proteinwas expressed in CHO cells after transtection.It may provide a direct and simplified method for primary assessment of the effect of TNFR1 siRNA on the(TNFR1) gene expression.

Objective To compare the treatment effect of calcium hydroxide and formocresol applied in root canal disinfection after root canal preparation.Methods 283 patients with 388 teeth diagnosed as chronic periapical were selected.Calcium hydroxide and formocresol were applied after opening of pulp chamber,pulp removed and root canal preparation.Calcium hydroxide was piped into the root canal by screw conveyor to fill the root canal in calcium hydroxide group,and formocresol cotton twist was setted in the root canal after dried in formocresol group.Tampons and ZOE were used for temporary closure.Treatment effects in both groups were compared in a week.Results The total effective rate in calcium hydroxide group was 96.39％,while 79.90％ in formocresol group.The total effective rote in calcium hydroxide group was higher than that in formocresol group(x2 =53.792,P ＜ 0.05).Conclusion Calcium hydroxide was safe,periapical tissue irritation and reliable effect was reached.It was worthy of clinical application.

This Paper reports the result of the differential Vi HAI test for typhoid carriers inXuzhou.One of 298 normal persons gave a false positive result.The traditional Vi HA I testfor 148 persons in the same group.6 revealed false positive.In 34 typhoid patients,thedifferential HAI test gave a titer of 1:12 to 1:64 in 16 cases(47%)Where as 10 of 239(4.1%)typhoid convalescents had the same titers.

Objective To understand the relevance of different coping styles and psychological reaction of the parents whose children with developmental dislocation of the hip (ODH). Methods To use symptom checklist (SCL-90) and simple coping style questionnaire, 96 parents of DDH children were questionnaired of the coping styles and psychological symptoms, and compared with the national norm, derived from the rating scale and the relationship between coping styles. Results Psychological symptom score of parents of DDH children was higher than the national norm, the psychological symptoms was objective, compared with the national norm, interpersonal sensitivity, depression, anxiety were statistically significant; interpersonal sensitivity, depression, anxiety was positively correlated with coping actively, somatization was positively correlated with the negative. Conclusions Parents of DDH children should be given more psychological counseling and psychological care,it may reduce the incidence of psychological symptoms,make them to take the right way to deal with.

Objective By measuring the concentrations of serum IL-2,IL-4,IL-8 and TNF-αin chronic bronchitis patients before and after the acupoint sticking therapy which was also called winter disease treated in summer to investigate the effect of therapy on serum concentrations of IL-2,IL-4,IL-8 and TNF-α.Methods 30 cases of patients diagnosed with chronic bronchitis in paracmasia and did not recieve other related treatment were randomly divided into acupoint application therapy group (treatment group)and the control group.The treatment group was giventhree Fuacupoint sticking therapy,the control group was given thecomfortacupoint sticking therapy.Serum IL-2,IL-4,IL-8 and TNF-αconcentrations were compared between the 2 groups before and after treatment.Results In the treatment group,the serum IL-2 concentration of patients after treatment was significantly higher than that before treatment(P <0.05),IL-8 and TNF-αconcentrations were significantly lower than those before therapy(P <0.05).Af-ter treatment,serum IL-2,IL-4 levels were significantly higher than those in the control group(P <0.05 )while IL-8 and TNF-αwere significantly lower than those of the control group(P <0.05 ).Conclusion Acupoint sticking therapy in summer for winter diseases effectively increase the concentration of serum IL-2 in patients with chronic bronchitis,reduced IL-8,TNF-α concentra-tions,and enhance their own immunity ability,reduce the degree of inflammation.

Physiologically based pharmacokinetic model (PBPK), a mechanistic mathematic model, which can simulate the absorption, distribution, metabolism and excretion of drugs, is being more widely used in pharmaceutical research and development areas. This article reviews primarily the recent advances in the procedure of establishing a PBPK model, including specifying of the PBPK model structure, specification of the tissue model, writing of equations, set of model parameters, simulation and evaluation. Application significance, major challenges and future developments of PBPK model in pharmaceutical areas are also discussed.

To explore the analgesic effect of propofol combine with dezocine after painless abortion and the influence on stress hormone and hemodynamics of the patients. Methods:Totally 126 cases of patients with painless abortion in our hospital from March 2013 to March 2014 were randomly divided into the observation group and the control group with 63 ones in each. The observa-tion group was anesthetized with propofol combine with dezocine, while the control group was anesthetized only by propofol. The dosage of propofol, anesthesia effect and postoperative recovery of the patients in the two groups were compared, and the level of stress hor-mones and hemodynamic parameters before and after the surgery were also compared. Results: Compared with that in the control group, the dosage of dezocine and the score of VAS after the surgery was significantly reduced in the observation group with obviously better anesthesia effect, and the differences had statistical significance (P<0. 05). The level of Cor, NE and ACTH was significantly improved, and the level of A-I and A-II was reduced significantly in the observation group, and compared with that in the control group, the differences showed statistical significance(P<0. 05). Compared with those that in the control group, the levels of SBP, DBP and SpO2 were improved significantly and the level of HR was reduced significantly in the observation group during and after the surgery, and the differences showed statistical significance (P<0. 05). The incidence of adverse drug reactions in the observation group was significantly lower than that in the control group, and the difference had statistical significance (P<0. 05). Conclusion:Propofol combine with dezocine for the surgery of painless abortion can reduce the dosage of dezocine, improve the anesthesia effect and reduce the harem pain with lower stress hormone fluctuation, hemodynamic effect and incidence of adverse reactions.

The relationships between cholinergic neurons and SP terminals were examined in the rat sacral ventral horn at the light and electron microscopic levels by means of double immunostaining methods. Cholinergic neurons were labeled by a monoclonal antibody to choline acetyltransferase (CHAT) with the avidin-biotin technique and stained bluish-green by indolyl-?-galactoside reaction products with ?-galactosidase as a marker. On the same sections, SP fibers were labeled by polyclonal antisera to SP after application of the peroxidase-antiperoxidase (PAP) method and stained brown by diaminobenzidine (DAB)reaction. At the light microscopic level CHAT-I neurons stained bluish-green and SP- I fibers Stained brown were found in the ventral horn. At the electron microscopic level, many asymmetrical axodendritic synapses(type I of Gray)were observed between CHAT-I dendrites and SP-I terminals in the ventral horn, but axosomatic synapses and symmetrical synapses (type I of Gray) were hardly detected. These results indicate that SP-I terminals make direct synapses with CHAT-I motoneurons of sacral ventral horn. These synapses may be predominantly excitatory and have importance in the control of muscular constriction.

Objective To explore the effect of case teaching method based on ADDIE model in the teaching of surgical nursing. Methods Two classes of the four-year nursing specialty college students (39 students in each class, 78 students in total) in the third grade were selected as samples. According to the principle of randomization, one class was set as the observation group, and the ADDIE model based on case teaching method was adopted. According to the five stages of ADDIE model, namely, analysis, design, development, implementation, and evaluation, the whole process was performed to achieve teaching goals and timely combination of clinical cases, so that the theoretical knowledge of teaching materials and clinical reality were highly connected. The other class was set as the control group. At the end of the semester, the teaching results were evaluated based on two groups of students' test scores and questionnaire findings. SPSS13.0 software was used for statistical analysis. t-test was used to test the significance of the test results,and chi square test was used to test the significance of the survey data. The results of the two teaching methods were compared with the control group. Results The scores of the two groups were significantly higher than those of the control group [(87.0±7.1) vs (78.0±8.6), P=0.000; (93.0±2.4) vs (85.0±3.8), P=0.000]. Significantly more students in the observation group (P<0.05) were satisfied with the teaching methods used to improve the enthusiasm and initiative of learning, improve their interest in learning, broaden their knowledge, improve the effect and efficiency of learning, inspire students to think, associate, and innovate, activate classroom atmosphere, improve the ability of students to analyze and solve problems, apply theories in practice, and accurately grasp the basic concepts and theoretical knowledge. Conclusion The case teaching method based on ADDIE model can effectively improve the teaching quality of nursing courses and improve students' ability to analyze and solve problems. It is well received by students, and it can enhance teachers' teaching skills as well.

Objective: To improve the training system of medical response to nuclear and radiological emergency, and strengthen the capability construction of medical response staff. Methods: By drawing lessons from the training management experiences on medical response of nuclear and radiological emergency of the developed countries and combining with the situation and status quo of China, to discuss how to improve the training system construction on medical response of nuclear and radiological emergency under the guidance of relevant polices. Results:Through researching, a series of countermeasures, such as standard organization system, improving contents and methods of training, strengthening the construction of teacher team, carrying out the evaluation for training effect and intensifying the medical response awareness for the whole people, were adopted based on the existent main problems on medical response training of nuclear and radiological emergency in China. Conclusion: The medical response training of nuclear and radiological emergency is a long term work involved with strong specialty and wide knowledge range. This work need constant innovation so as to strengthen the training system construction of medical response, and train high competent talents on medical response of nuclear and radiological emergency, and then to establish a omnibearing, multilateral, scientific, systematic and long-acting training mode.