Objective To express,purify and identify human monomolecular antibody against Clostridium perfringens type A α-toxin,in order to lay a foundation for the prevention and treatment of various diseases caused by this toxin.Methods The fully human single-chain fragment variable(ScFv) gene against Clostridium perfringens type A was linked with the constant region of light chain and heavy chain of human antibody in different combinations to construct multiple monomolecular antibody expression plasmids against Clostridium perfringens α-toxin,which were expressed in competent E.coLi BL21(DE3).Indirect ELISA was used to detect the immunobinding activity of the monomolecular antibodies,and the monomolecular antibody with the highest immunobinding activity was purified by SepharoSe 4 FF and rProtein-A FF affinity chromatography,The purified products were analyzed by 12% SDS-PAGE.Indirect ELISA was used to detect the immune binding activity of each monomolecular antibody.Results Five recombinant plasmids,PTS-ScFv-CL-CH_2-CH_3,PTS-ScFv-CH_2-CH_3,PTS-ScFv-CL-CH_2,PTS-ScFvCH_2,and PTS-ScFv-CL,were constructed.After transfection into E.coli BL21(DE3) and purification,the corresponding monomolecular antibodies,ScFv-CL-CH_2-CH_3,ScFv-CH_2-CH_3,ScFv-CL-CH_2,ScFv-CH_2,and ScFv-CL,were obtained,which had the relative molecular mass of about 60 000,and the concentrations of about 1 mg/mL.Among them,ScFv-CH_2-CH_3showed the highest immune binding activity,and the A_(450) value reached 3.9,much higher than the other four monomolecular antibodies,with the concentration of about 1 mg/mL and the purity about 86%.Conclusion A fully human monomolecular antibody ScFv-CH_2-CH_3 with high affinity,low immunogenicity and internalization activity was obtained,which lays a foundation for the further study of therapeutic antibody against CPA.

Objective To explore the effect of Tracer Methodology on the physical restraint management of the elderly patients. Methods The idea of Tracer Methodology was based on a case study of physical restraint, and then complementary tracking and system tracking. The key problem, which was founded in using Tracer Methodology on the physical restraint management of the elderly patients, could be continuously improved from the regulations, procedures, training, implementation, inspection, feedback, rectification, the implementation and the effectiveness. The normative rate of physical restraint was compared before and after the implementation of this method. Results After using the Tracer Methodology, the rate of physical constraint in the elderly patients was 82% (41/50), which was significantly increased (χ2=5.877,P=0.015) compared with before implementation of this method, which was 60% (30/50). Conclusions Tracer Methodology could improve the normative rate of physical restraint in elderly patients. Meanwhile, it was beneficial for nurses to participate in quality management, and strengthen the team cooperation.

Action Potentials ; drug effects ; physiology ; Animals ; Cadmium ; toxicity ; Calcium Channels, L-Type ; physiology ; Myocytes, Cardiac ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley

Objective To investigate the pressures from night shifts among nurses from pediatrics department and find the countermeasures.Methods A self-designed questionnaire was used to investigate the pressures from night shifts among 112 nurses from pediatrics department as well as the reasons.Results 110 of the nurses(98.2%) assumed that the pressures from night shifts was intensive.The main reasons for the pressures came out of heavy workload,shortage of staff and stuff and the unsettled balance between job and family.Conclusion Such measures as reasonable distributions of nurses,abound supply of stuff and reduced working links may be effective for the alleviation of the pressures on nurses at night shifts,motivated activeness in them and then the enhancement of nursing quality.

Objective To evaluate the profile of transient ischemic attacks in younger patients compared with older patients effectively.Methods We study 75 younger patients(≤45 years)compared with 90 older patients(≥65 years).History of presenting transient ischemic attacks,etiology,clinical feature and lab investigation were compared on the basis of the above age groups.Results Overweight、hypercholesterolemia、insomnia and vascular disease family history being more common in the younger group and Hypertension,ischemic heart disease,long-term history of smoking being more common in the elder group.Diabetes and high serum uric acid occurred high frequently in both groups.Conclusion A significant correlation existed between etiologies contribution and adverse life custom,the early management should be take up to prevent TIA occurrence.

Objective To evaluate the effects of dexmedetomidine on the hemodynamics and the heart rate variability(HRV)in patients under-going lower limbs operations with application of tourniquet. Methods Forty patients undergoing lower limbs operations with application of tourni-quet were randomized assigned to dexmedetomidine group(group D,n=20)or control group(group C,n=20). After combined spinal-epidural anesthesia,group D received a continuous infusion of dexmedetomidine(0.5μg/kg for 10 min for loading dose and followed by 0.2μg·kg-1·h-1) until tourniquet deflation. The control group received normal saline instead. Mean arterial pressure(MAP),heart rate(HR),saturation of pulse ox-imetry(SpO2),low frequency power(LF),high frequency power(HF)and LF to HF ratio(LF/HF)were recorded at regular time points:imme-diately before loading dose(T0),before tourniquet inflation(T1),15 min after tourniquet inflation(T2),30 min after tourniquet inflation(T3),45 min after tourniquet inflation(T4),60 min after tourniquet inflation(T5),1 min after tourniquet deflation(T6),5 min after tourniquet deflation (T7)and 10 min after tourniquet deflation(T8). Results Compared with T0,the MAP of group D significantly decreased at T6-T8(P<0.05). Compared with T0 and T1,the MAP of group C increased at T2-T5(P>0.05). Compared with T2-T5,the MAP of group C significantly decreased at T6(P<0.05). Compared with group C,the MAP of group D significantly decreased at T6 and T7(P<0.05). Compared with T0,the HR of group D significantly decreased at T1-T8(P<0.05). Compared with T0,the HR of group C had no significant change at T1-T5(P<0.05). Compared with T1-T5,the HR of group D and group C significantly increased at T6(P<0.05). Compared with group C,the HR of group D significantly decreased at T1-T4 and T6(P<0.05). Compared with T0,the SpO2 of group D and group C significantly decreased at T6(P<0.05). Compared with group C,the SpO2 of group D significantly decreased at T1-T3(P<0.05). Compared with T6,LF of group D and group C significantly increased at T7(P<0.05). LF were comparable between groups D and C(P>0.05). Compared with T0,the HF of group D significantly increased and the LF/HF of group D significantly decreased at T1-T4(P<0.05). Compared with group C,the HF of group D significantly increased and the LF/HF of group D significantly decreased at T1-T4(P<0.05). Conclusion The appropriate dose of dexmedetomidine(loading dose 0.5μg/kg and maintenance dose 0.2μg · kg-1 · h-1)can significantly increase vagal tone and improve cardiac sympathetic and parasympathetic balance during tourniquet appli-cation.

Objective To evaluate the influence of black dots,isolated line above the best correct visual acuity and multiple lines containing the best correct visual acuity in the accuracy of Jackson cross cylinder examination.Methods Forty students were chosen(11 males,29 females,between 24 to 29 years old).Routine examination of phoropter was used to check under the maximum plus to maximum visual acuity(MPMVA) status.The sequence of three fixation targets was chosen randomly.Astigmatism chart was used to judge the best one when the different results were obtained.Results Among the 80 eyes of 40 students,12 eyes were spherical(the cylinder degree below 0.25 DC).The response of three fixation targets was the same in 19 eyes(27.9%),23 eyes(33.8%)were sensitive to the black dots target,14 eyes(20.6%)were sensitive to the isolated line above the best correct visual acuity,12 eyes(17.6%) were sensitive to multiple lines containing the best correct visual acuity.Kruska Wallis Test was used to detect the difference among three groups.No significant difference was found(?2 =4.026,P=0.134).Conclusion According to the efficiency of examination,communication and judgment,black dots target is recommended although no difference is detected among three fixation targets.

Objective To explore the impact of physical activities on adolescents' external problem behaviors and the mediating effect of self-esteem.Method The physical activity rating scale(PARS-3),Achenbach adolescent self rating scale,Rosenberg self-esteem scale and adolescent physical self-esteem scale were used to investigate 419 participants selected from middle school students in Beijing.Results Adolescents who participated in physical activities with moderate amount showed less external problem behaviors.Adolescents who participated in physical activities with large amount appeared to be of more self-esteem.Both self-esteem and body-esteem showed partially mediating effects between physical activities and disciplinary offence.Conclusion Adolescents conducting moderate amount of physical activities can enhance their self-esteem,and ultimately reduce the incidence of their external problem behaiviors.

Ureaplasma urealyticum(UU)is one of the most common pathogen in childbearing age women.The proportion of neonate especially premature baby infected with UU is increasing yearly.UU infaction is related with premature labour,low birth weight,bronchopulmonary dysplasia,chronic lung disease,respiratory distress syndrome and other perinatal diseases by promoted the expression of inflammatory cytokines,increasing the inflammatory response and interfering inflammation clear.There still has controversial point to treat perinatal diseases caused by UU infection by erythromycin,azithromycin,pulmonary surfactant,steroid.

To observe the clinical efficacy of Artisan iris-clip intraocular lens ( lOL) implantation on cataract with a wide range of lens dislocation.
●METHODS: Twenty-four hospital patients (28 eyes) of cataract with lens subluxation by trauma (20 patients, 20 eyes) and Marfan's syndrome (4 patients, 8 eyes) during May 2007 to December 2011 were selected. All of them underwent phacoemulsification or an anterior vitrectomy and Artisan iris - clip lOL implantation at stage Ⅰ. Postoperative visual acuity, corneal endothelial cell count, intraocular lens position and postoperative complications were observed. Follow-up time was 6mo.
●RESULTS: Compared with the preoperative, postoperative best corrected visual acuity improved to varying degrees, visual acuity of 0. 1-0. 5 was in 20 eyes, 0. 5-1. 0 in 8 eyes. After followed up 1-6mo, no artificial lens shift, iris inflammation, iris atrophy and corneal decompensation and other complications were observed.
●CONCLUSlON: Cataract surgery with a wide range of lens dislocation is difficult, but as long as surgeons who have a wealth of clinical experience and superb skills, still can carry out phacoemulsification surgery or anterior vitrectomy and Artisan iris - clip lOL implantation and obtain a better outcome.