Objective To establish a method for quantitative detection of total immunoglobulin E(TIgE) by Time resolved Flu‐oroimmunoassay(TRFIA ) .Methods The method for quantitative detection of TIgE by TRFIA was established on the basis of solidphase double sandwich enzyme linked immunosorbent assay(ELISA) .The methodology was evaluated .Results The TIgE TR‐FIA intra assay and inter assay coefficients of variation (CV) were 1 .59% -1 .68% and 5 .23% -7 .33% ,respectively .The lower limit of detection was 0 .25 IU/mL .The linear range was 1 .47-1 510 .00 IU/mL .The accuracy was within the allowable deviation ( ± 10% ) .The recovery rate was 97 .00% -106 .75% .The cross reaction test and interference experiment could meet the testing requirements .The TIgE TRFIA showed no HOOK effect at least up to 15 000 IU/mL TIgE ,compared with EUROIMMUN ELISA ,the correlation coefficient(r) was 0 .999 2(P<0 .01)for 40 blood specimens in the range of 14 .43-518 .81 IU/mL ,and the expected bias was in the range of acceptable bias (± 12 .50% ) .The reference value 100 IU/mL could be used for a normal ,allergy free adult sample TIgE level detected by TRFIA .Conclusion The established TRFIA for TIgE detection meets the demand of clini‐cal application with good precision ,high sensitivity ,wide linear range ,high accuracy ,specificity and other advantages .

Objective To investigate the role of molecular allergen diagnosis in respiratory allergic diseases, and measure the prevalence and the allergen molecular mapping of the asthma and allergic rhinitic patients.Methods Totally 80 cases of asthma or allergic rhinitis patients with two or more inhaled allergens from First Affiliated Hospital of Guangzhou Medical University during 2013 January to 2014 December were tested for serum specific immunoglobulin E ( sIgE ) against 112 allergen components by ImmunoCAP ISAC.Results About 78.8% ( 63/80 ) patients is three or more allergen components sensitization, the most prevalence allergen components were house dust mites′Der f 1 67.5%(54/80), Der f 2 66.2%(53/80), Der p 1 and Der p 2 were 63.8%(51/80) and 61.2%(49/80), Der p 10 was only 12.5%(10/80), follewed by Fel d 1 22.5%(18/80), Cyn d 1 16.2%(13/80), Phl p 4 15.0%(12/80), Pen m 1,Pla a 2,Jug r 2 and Can f 1were all 11.2%(9/80), Lep d 2, Blo t 5, Bla g 7 and Ani s 3 10.0%(8/80).Other allergen components positive rates were low.Conclusions Molecular allergen diagnosis can provide more comprehensive diagnostic information for allergic patients, by providing allergen map.Allergen microarray can discover the neglected allergens such as bermuda grass, timothy grass and plane tree pollen allergens in southen China.

Objective To investigate the types and distributions of allergens, and the responsiveness to these allergens SIgE and tIgE as related to allergic disorders in patients with allergies in Guangdong. Methods Serum samples were obtained from patients with allergic disorders (n = 7 144) who visited our hospital between 2009 and 2014. The sera were subjected to analysis of 15 common allergens. Results The positive rates of sIgE and tIgE were 62.4%and 54.6%, respectively. Dermatophagoides pteronyssinus(Der p) had the highest prevalence of aeroallergen-specific IgE and milk the highest prevalence of food allergen-specific IgE. Other aeroallergens and food allergens produced mild responses except Der p and Der f. The sensitization peak of Der p and Der f appeared at the age of 10 to 12 years. The sensitization peak of milk appeared at the age of less than 3 years and that of eggs did at the aged of 4 to 6 years. The averaged tIgE positive rate went up with the increase in the number of allergen sensitization. Conclusion Der p, Der f, milk and eggs are major sensitizers responsible for common allergic disorders in Guangdong. Knowledge concerning allergen characteristics at various age groups may be helpful for early diagnosis and intervention for allergies.

Objective To explore the positive distribution characteristics and analyse the correlation of common food allergen-specific immunoglobulin E (sIgE) with suspected food allergy in the First Affiliated Hospital of Guangzhou Medical University.Methods Using fluorescence enzyme-linked immunosorbent assay to detect the serum sIgE antibody of 854 patients,including 7 kinds of food allergens (milk,egg white,egg yolk,shrimp,crab,peanut and soybean) from July 2006 to January 2013.Data were processed by statistical software SPSS19.0.Results The positive rates of the 7 kinds of food allergen were 39.3％ (283/720),36.3％ (216/595),9.8％ (28/285),21.2％ (36/170),24.3％ (17/70),14.8％ (9/61) and 10.0％ (5/50).Two hundred and eighty-two patients were detected egg white and egg yolk sIgE,the positive rate of egg white sIgE 59.6％ (168/282) was much higher than the egg yolk sIgE 9.2％ (26/282).The positive rate of the milk and egg white sIgE were different between the age groups and decreased with the age(milk:x2 =792.88;egg white:x2 =658.21,P ＜ 0.01).The degree in level 4 or above of milk and egg white sIgE were 1.08％ (2/186) and 1.12％ (2/178) in positive patients.Five hundred and twenty-eight patients were detected milk and egg white sIgE,26.9％ (142/528) were both positive,sIgE levels in serum of them was moderately correlated (rs =0.758,P ＜ 0.01).Serum sIgE of shrimp and crab was detected in 64 patients,16 cases was positive of the shrimp,17 cases of the crab was positive,both positive in 16 cases,sIgE levels in serum of them were highly correlated (rs =0.973,P ＜0.01).In the simultaneous detection of 34 patients' serum sIgE of peanut and soybean,peanut was positive in 4 cases,soybean was positive in 3 cases,both positive in 2 cases,sIgE levels in serum of them were highly correlated (rs =0.879,P ＜ 0.01).Conclusions Egg white is the major allergen of egg allergy.The correlations of shrimp and crab,peanut and soybean are extremely high,probably because of their homology; egg white and milk are not homologous food,but the levels of sIgE between them display a moderately correlation,indicating the existing of synergistic reaction.

Objective To analyse distribution of age and gender characteristics of specific IgG(sIgG)antibodies and specific IgE (sIgE)antibodies of 13 types of food allergens in patients with food anaphylaxis,and to explore the relationship between sIgG and sIgE in food anaphylaxis.Methods 314 cases of patients from 2009 to 2012 were selected as subjects,and divided into underage group(1 63 cases)and adult group(1 5 1 cases).Serum sIgG of 13 types of food allergens were detected by using enzyme linked immu-nosorbent assay,serum sIgE of these food allergens were detected by using immune capture.Results 80.25% of the patients were sIgG-positive,and no obvious gender differences were found;while the positive rates of sIgG in the underage group(94.48%)were higher than that in the adult group(64.90%),there were statistically significant differences(P < 0.05 ).34.39% of the patients were sIgE-positive.The positive rates of sIgE in male patients(40.68%)were higher than that in female patients(26.28%),and that in the underage group(55.21%)were also higher than that in the adults group(1 1.92%),there were statistically significant differences(P <0.05).Conclusion The total positive rates and its distribution characteristics of sIgG and sIgE of same food aller-gens were obviously different.Food anaphylaxis might be associated with age,gender,food types and individual diversity.

Objective:To find related polypeptides which could bind interleukin (IL) 5 with high affinity.Methods:The recombinant human IL 5 was biotinylated with NSH LC Biotin,then biotinylated IL 5 was used to react with random peptide library displaying 7 amino acids fused on protein III of M13 Phage for three rounds biopanning.The selected clones were assayed by sandwich enzyme linked immunosorbent assay(ELISA)and competition ELISA.The positive clones with high affinity were used for automated sequencing with dye labeled dideoxynucleotides,and the amino acid sequence of polypeptide displayed on phage was deduced.Results:The enrichment was shown by ELISA after 3 rounds of biopanning.9 positive clones could bind to IL 5 with high affinity.Sequencing of the genes encoding these peptides on 9 positive clones showed some conserved epitope information such as SX 1 2 AS,ALAS.Conclusion:Potential polypeptides binding IL 5 with high affinity could be selected from phage display peptide library,SX 1 2 AS may be the motifs recognized by IL 5.

Objective To discuss the clinical effect of different surgery in the treatment of shunt-dependent hydrocephalus by comparing the postoperative complications of ventriculoperitoneal(V-P) shunt surgery with different incidence of eminence puncture or the triangle puncture.Methods 98 patients with shunt-dependent hydrocephalus who underwent V-P shunt were analyzed.According to different operation methods,they were divided into eminence puncture group (63 cases) and triangle puncture group (35 cases).The operation time and postoperative complications were compared between the two groups,and the clinical efficacy was compared by Glasgow Coma Scale (GCS) score at the end of 12 months follow-up.Results The operation time of the eminence puncture group [(53.35 ± 5.72) rain] was longer than (35.52 ± 9.18) min of the the triangle puncture group (t =6.024,P ＜ 0.05).The incidence rate of complication in the eminence puncture group was lower than that in the the triangle puncture group at postoperative 12 months (x2 =4.395,P ＜ 0.05).Postoperative CT examination found that the ventricle width of the two groups was decreased in different degree.The improvement of Evan's index(t =40.86,P ＜0.05) and GCS score(U =4.50,P ＜ 0.05) in preoperative of the eminence puncture group were greater than those of the triangle puncture group after 12 months.Conclusion The clinical efficacy of V-P shunt of the frontal lateral ventricle puncture may be superior to the triangle puncture in the treatment of shunt-dependent hydrocephalus.

The article introduces the training of basic skill,renovating of the experiment contents,the diversifi cation and abundance in interest of experiment teaching methods of the students of medical laboratory speciality of higher profession.

Cultivation of laboratory physicians in accordance with the needs of discipline should be a priority of education on laboratory medicine in China. Unfortunately, the professional career of Chinese laboratory physicians is presently entrapped in a dilemma owing to unfavorable settings, in-cluding the late start of laboratory physician training in this country, unmatched installation between higher school faculty and curriculum, faulty programs of in-hospital training, disparity in management policies, low social esteem, uncertainty in personal intention of employment, and poor knowledge of position awareness. This paper puts forward a lot of measures such as the development trend of the university curriculum, which is in compliance with the subdivision and refinement of clinical medicine, the establishment of a medical degree for a professional laboratory physician, constantly improving the standardization of laboratory physician residency training and continuing education system for the in-spection of doctors , setting up physicians inspection posts and giving a clear mandate to examine physicians inside the hospital, while outside of the hospital continually improving awareness and accep-tance of the laboratory physicians, giving more educational resources to laboratory physicians, so as to improve the overall level of the laboratory medicine.

Objective To observe the pathology in temporal lobe cortexes and hippocampus in protopathic intractable temporal lobe epileptics.Methods The cortexes of spike foci in temporal lobe and hippocampus were obtained from 9 cases with protopathic intractable temporal lobe epilepsy who accepted operation.The samples were observed under the transmission electron microscope.Results The ultrastructure changes in spike focus of temporal lobe cortexes are similar to those in hippocampus.It is common that neurons were pycnotic and decreased.Astrocytes were hydropic and degenerative.Gliosis were found in some cases.The number of synapses increased or decreased in different cases and positions.Blood-brain barriers were destroyed because foot processes of astrocytes around capillaries were edematous.Conclusion Neuronal loss,gliosis and synaptic reorganization which occur in epileptic hippocampus and maybe also in epileptic temporal lobe cortexes destroy the balance between excitatory and inhibitory neurotransmission.The abnormalities probably associate with protopathic intractable temporal lobe epileptic seizures.