Objective To develop an indirect ELISA-based peptide scanning method combined with nuclear magnetic resonance(NMR)technique for the epitope identification of calcitonin gene-related peptide(CGRP)antibodies.Methods The antigen binding activities of two antibodies(new CGRP antibody and control antibody)were determined by indirect ELISA using each truncated CGRP fragment as coating antigen,and the linear epitope was analyzed according to the EC50value of four-parameter curve. Two-dimensional hydrogen-nitrogen correlation(2D1H-15N HSQC)spectrum of CGRP were acquired by NMR technique,and the binding of antibodies to the arginine of CGRP were analyzed through the disturbance of the antibodies to CGRP signals. Specific arginine modifications were detected by liquid chromatography-mass spectrum(LCMS) and NMR technique,and two arginine resonances were assigned on CGRP by correlating the rank order of the modification rate.ResultsThe antigen binding activities of two antibodies with CGRP(1-37),CGRP(19-37)and CGRP(25-37)showed dose-response relationships,and were fitted with four-parameter equation. However,there were no significant antigen binding with CGRP(1-18),CGRP(19-24)and CGRP(25-37)without C-terminal amide. The linear epitopes of both antibodies were located at the C-terminal of CGRP. The resonances of arginine ε-NH in 2D1H-15N HSQC spectrum disappeared in the presence of the control antibody;and the resonances appeared in the presence of the new antibody. The arginine R11 and R18 of CGRP could bind to the control antibody,but not to the new antibody. The NMR assignment for the arginine resonances were made by correlating the relative ranking of the modification rate where signals A and B arose from R11 ε-NH and R18 ε-NH respectively.ConclusionIn this study,the linear and conformational epitopes of new CGRP antibody and control antibody were identified based on the methods of ELISA and NMR,which may provide a theoretical basis for the design of the candidate therapeutic CGRP antibodies.

Objective To analyze the host cell protein(HCP) residues in bevacizumab injection and its downstream productsby nano ultra-performance liquid chromatography-tandem mass spectrometry(Nano-UPLC-MS/MS) and enzyme-linked immu-nosorbent assay(ELISA).MethodsThe samples were directly treated with trypsin, and the undigested antibodies wereprecipitated by heat treatment, which were then detected by Nano-UPLC connected with Q-Exactive Mass Spectrometer. Thecollected peptide information was compared with the total protein library of Chinese hamsters by MaxQuant software, and theHCP types were identified. The absorbance values of samples were detected by double-antibody sandwich ELISA, and theHCP content was calculated according to the four-parameter curve.ResultsAfter purification by downstream process, thelevels and types of HCPs decreased significantly. The content of HCPs in injection was 7 ppm, including 20 types, amongwhich 5 types of persistent residual HCPs were detected, and 4 types of high-risk HCPs were not detected. Compared withthe original drug, the HCP residues of the two drugs were both less than or equal to 15 ppm, and there were differences inHCP types due to different purification processes.ConclusionThe combination of Nano-UPLC-MS/MS and ELISA providesa comprehensive analytical strategy for the characterization and quantification of HCP, especially the types and distributionof persistent residual HCP and high-risk HCP, which can provide a reference for the process optimization, comparabilityresearch and safety risk assessment of products.

Objective To investigate the potential factors that may be associated with clinical significant endoscopy findings (CSEF) and the characters of the appropriate patients for upper endoscopy.Methods Information of the out-patients was collected from the questionnaires that were performed before undergoing upper endoscopy from 26 September 2011 to 23 December 2011,including demographics characteristics,symptoms,gastroesophageal reflux disease questionnaire (Gerd Q) score,comorbidities,medication and purpose for upper endoscopy.The analyses were performed by Logistic regression to find the potential factors that were associated with CSEF.Results A total of 942 cases were enrolled.There were 471 (50.0％) patients with dyspepsia and reflux symptoms,300(31.8％)patients with dyspepsia and without reflux symptoms,and 86(9.1％)patients with reflux symptoms and without dyspepsia.Sum to 325 (34.5％) patients were diagnosed with CSEF,119(12.6％) with erosive esophagitis,28(3.0％) with Barrett esophagus,102 (10.8％)with peptic ulcers,66 (7.0％) with gastric dysplasia,and 13 (1.4％) with upper malignancy.Multivariate Logistic regression analysis showed that men (OR=1.677,95％CI:1.148 to 2.451),older age (OR=1.032,95％CI:1.021 to 1.044),alcohol intake (OR=1.761,95％CI:1.068 to 2.903),Gerd-Q score increase (OR=1.079,95％ CI:1.003 to 1.160),and presence of acid regurgitation (OR=1.659,95 ％CI:1.143 to 2.408) were significantly associated with increasing risk of diagnosis for CSEF,while taking proton pump inhibitors (OR=0.298,95％CI 0.109 to 0.818)was associated with lower possibility of detection.Conclusions Factors such as male,older age,alcohol intake,Gerd-Q score increase,presence of acid regurgitation and taking proton pump inhibitors,may have a certain significance for screening out patients and further improving the cost efficiency of gastroscopy.

Objective To survey the incidence of depression in Parkinson disease(PDD) and explore the related factors of PDD.Methods 127 patients with Parkinson disease were investigated,all of them were assessed by Hamilton Depression Rating Scale,self-designed register form and the Unified Parkinson disease scale(UPDRS).A stepwise multiple factor regression analysis was carried out to determine factors that best accounted for the occurrence of PDD.Results The overall incidence rate was 47.2%.The minor,moderate and major incidence was 14.2%,28.3% and 4.7% respectively.The regression analysis showed that factors accounted for the occurrence of PDD were gender,the severity of PD,stage of patient's motor impairment,education level and the use of dopamine agonists.Conclusion PDD seems to have a multifactorial etiology.Knowledge of risk factors related to the occurrence of PDD may help guide neurologists in the early prediction and treatment of this disease.

Objective To investigate the duration of the first-fundus-examination and the status of diabetic retinopathy in patients with type 2 diabetes mellitus. Design Questionnaire. Participants 956 patients (1905 eyes) with type 2 diabetes in Tongren Eye Center of Beijing Tongren Hospital. Method Self-designed questionnaire was carried out. Patients were divided into different groups by the du-ration of the first-fundus-examination, degree of diabetic retinopathy, duration of diabetes, education background and living area in the data analysis. Main Outcome Measurements Duration of the first- fundus-examination, degree of diabetic retinopathy. Results Dia-betes patients in shorter duration groups finished the first-fundus-examination earlier than in longer duration groups (all P <0.05). Dif-ferent education background didn't show significant influence on duration of the first fundus examination. Urban patients received earli-er first-fundus-examination than rural patients (all P <0.05). The incidences of vitreous hemorrhage, retinal detachment and retinal pho-tocoagulation were rising with the duration of diabetes. The proportions of vitreous hemorrhage and retinal photocoagulation were higher in primary-school education group than in middle-school and college education groups (all P<0.05). Difference of the incidences of vit-reous hemorrhage and retinal detachment weren't found between in Beijing and non-Beijing patients. The incidences of vitreous hemor-rhage and retinal detachment in urban patients were higher than in rural patients (all P<0.05). Conclusion The most diabetic patients finish the first-fundus-examination much later than requested. Diabetic patients in lower-education-level groups and in rural group are even later, and their conditions are more serious.(Ophthalmol CHN, 2009, 18: 279-282)

Objective To evaluate correlation of CT features of tuberculous adrenal glands with duration of adrenal insufficiency and propose a new staging criteria for adrenal TB Methods CT features of 28 cases of documented adrenal tuberculosis were analysed retrospectively The patients were divided into 3 groups according to the duration of adrenal insufficiency: group A (10 cases , less than 1 year), group B (13 cases, 1 year to 4 years), group C (5 cases, longer than 4 years) The main CT features analyzed included size and shape of the adrenal gland, calcification ,and low density foci Results Enlargement of the adrenal gland: group A(10) , group B (13), group C (1) Basic configuration of adrenal gland were identifiable in group A ,whereas others had more irregular shape Calcification: group A (2) , group B (11),group C (5) Low density foci: group A (3), group B(0), group C (0) Conclusion CT features of adrenal glands correlated with duration of adrenal insufficiency in adrenal tuberculosis It is more reasonable to use tri phase criteria according to CT appearance in adrenal tuberculosis

Objective To estimate the expressions of cyclin D1 and Ki67 in esophageal squamous cell carcinomas and their clinical significance. Methods The expressions of cyclin D1 and Ki67 were detected by immunohistochemistry in 28 specimens esophageal squamous cell carcinoma (ESCC) and 33 samples esophagitis tissue. Results The positive expression rate of cyclin D1 in ESCC and esophagitis tissues was 60.7% (17/28) and 33.3% (11/33) respectively (?2=4.573,P=0.032),while the mean Ki67 label index (Ki67 LI) was (49.21?25.15)% and (11.62?9.87)% respectively (t=7.908,P=0.000). The positive expression rate of cyclin D1 in TNM stage I tissues was 14.3% (1/7),which was significantly lower than that in stages Ⅲ and Ⅳ tissues[85.7% (6/7) and 100% (5/5),P=0.029 and 0.015,respectively]. In cases with lymph node metastasis,the positive rate of cyclin D1 expression was 90.9% (10/11) that was significantly higher than that in those without lymph node metastasis (P=0.016). The 28 patients with ESCC were followed up for 6 to 34 months [mean,(25.0?4.2) months],during which 19 patients survived,4 patients died of deterioration of the primary diseases (3 cases) or cerebrovascular affair (1 case),and the other 5 patients was lost to follow-up. Conclusions The expression of cyclin D1 is correlated with advanced tumor and lymph node metastasis. The Ki67 is highly expressed in ESCC tissues; no relationship exists between the level of the expression and the pathological differentiation type and TNM stage of ESCC.

The Na+/H+ antiporter in vacuolar membranes transports Na+ from the cytoplasm to vacuoles using a pH gradient generated by proton pumps, which could reduce Na+ toxicity. It is uncertain that whether the woody plants have the same mechanism. Through differential centrifugation and sucrose density gradient centrifugation, tonoplast vesicles were isolated from Populus tremula calli broken by blender. After establishing pH gradient by V-ATPase, Na+ could dissipate the pH gradient, which indicates that there is Na+/H+ antiporter in the tonoplast vesicles from Populus tremula calli (Km=11.4 mmol/L). Amiloride could inhibit the Na+/H+ antiporter activity. The antiporter could transport Na+ and K+, the affinity for Na+ is higher. Salt stress decreased Km and Vmax.

Objective To investigate the clinical efficacy of topical corticosteroids combined with fusidic acid cream in treating infantile eczema.Methods Four hundreds cases of infantile eczema in the outpatient department of our hospital from March 2012 to January 2015 were collected and divided into observation group (220 cases) and control group (180 cases).The observation group received topical corticosteroids combined with fusidic acid cream,while the control group was treated only by topical corticosteroid.The medication time reaching to clinical cure,effect maintenance time during 30 d observation period and recurrence rate were recorded in the two groups.Results During the 30 days of observation, 50 cases of infantile eczema withdrew from the study,17 cases in the observation group and 33 cases in the control group.The average medication time in the observation group was (2.2±0.9)d,which was shorter than (3.2±1.1)d in the control group.The effect maintenance time during observation period in the observation group was (11.7±5.4) d,which was longer than (7.2±4.0)d in the control group,the difference was statistically significant(P<0.05);74.0% of recurrent cases during the observation period in the observation group manifested by mild eczema,while 57.8% in the control group manifested by mild eczema and had no need to use corticosteroid,the improvement of symptoms during recurrent period for the patients in the observation group was better than that for the patients in the control group,the difference was statistically significant (P<0.05).Conclusion Topical corticosteroids combined with fusidic acid cream for treating infantile eczema can reduce the medication time,prolong the effect maintenance time,and alleviate the recurrent symptoms.

Objective:To understand the relationship between psychiatric nurses' communication ability, personality characteristics and conflict management patterns, and to provide a basis for training psychiatric nurses' conflict management ability courses.Methods:A convenient sampling method was used to investigate and investigate 208 nurses in the first-line clinical hospitals of a third-class psychiatric hospital, used the nurses′ clinical communication ability scale, the nurse′s interpersonal conflict management morphology scale, and the simplified version of the Chinese Big Five Personality Questionnaire.Results:Integration dimension (18.67±2.58) points, concession dimension (12.93±2.43) points, escape dimension (14.05±2.02) points, dominance dimension (14.49±3.06) points, and integration dimension and gender ( t value was -2.57, P=0.01)), education ( F value was 3.76, P=0.02), technical title ( F value was 3.03, P=0.02), position ( t value was 2.08, P=0.03), whether to participate in nurse-patient communication training ( t value was 2.54, P=0.02). Nurses with pleasant personality could better deal with interpersonal conflict ( r=0.361, P<0.01), and the nurse′s personality had a direct effect on the form of interpersonal conflict ( P=0.036, 95% CI 0.107-0.422), the nurse′s communication ability plays a part of indirect effect ( P=0.031, 95% CI 0.001-0.063). Conclusion:The form of interpersonal conflict management of psychiatric nurses was closely related to communication ability and personality characteristics. Nursing managers could carry out targeted training to improve the conflict management ability of nursing staff.