Quantitative immunoassay technique is the common method of quantitative detection in clinical laboratory. Several important branches of quantitative immunoassay were formed by changing the tracer or the Antigen-antibody complex separation method, including radioimmunoassay, fluorescent immunoassay, enzyme immunoassay, chemiluminescence, colloidal gold, immuno-turbidimetric analysis and homogeneous immunoassay. Different immunoassay techniques have their own characteristics, also apply to different detecting conditions in clinic. This paper reviewed several common kinds of quantitative immunoassay technology, and discussed both their advantages and disadvantages, which provide reference for the application and development of clinical testing technology.

Assay"'of Epstein-Barr virus (EBV) DNA was conducted in the tissues of 60 patients with nasopharyngeal carcinoma (NPC), 30 patients with chronic inflamation of nasopharyngeal mucous membrane and 40 patients with other malignant epithelial tumor respectively using DNA in situ hybridization method. The positive rate of EBV-DNA in patients of NPC was 71.6%, while it was negative in pat ents with other malignant tumors a small number of EB-VDNA positive cells were also discovered in the epithelial cells of paratumours and chronic inflamation of nasopharyngeal mucous membrane. Significant correlation was found in the study of serum EBV-antiby in patients of NPC and EBV-DNA in cancer cells. The results suggest that EBV may be of etiologic significance in the pathogenesis of NPC.

Objective To gain the virus like particles (VLPs) based on Trop2 targets ,which provided the basis for further in vi‐vo induced experiments and anti‐tumor vaccine studies .Methods Using molecular cloning method to constract eukaryotic expres‐sion vector of pCAGGs/Trop2 and baculovirus expression vector of pFastbac1/Trop2 ,expression product of pCAGGs/Trop2 in HeLa cells were intended to be to be anchored to the cell membrane by the methods of Immunohistochemistry ;pFastbac1/Trop2 were transformed into E .coliDH10bac isolates to gain recombinant bacmids ,which were transfected into insect cells to express re‐combinant baculovirus with Gag rBV ,then sucrose density gradient centrifugation ,Western Blot and electron microscope were per‐formed to purify and identify the Trop2 VLPs .Results Building recombinant bacmids which were transfected into insect cells to express recombinant baculovirus with Gag rBV ,gained the recombinant Virus like Particles .Conclusion Trop2 VLPs was success‐fully prepared ,which laid the foundation for the subsequent induction of humoral and cellular immune response .

OBJECTIVE: By analysing the video-nystagmography findings of positional tests,to evaluate the therapeutic effect of the patients with horikontal semicircular canal cupulolithiasis (HSC-Cup). METHOD: A retrospective study of 36 patients with HSC-Cup. The induced nystagmus in roll tests was recorded by videonystagmography, whose direction, latency, intensity and time characteristics were analysed. All of the 36 patients were treated with lying position avoiding normal side and oral-taken betahistine mesilate tablets. A week later return visits and curative effects evaluation were made. RESULT: Horizontal apogeotropic nystagmus was induced by turning left or right in HSC-Cup roll tests. The time of latency and duration turning to normal and lesion side were(0. 93 ± 0. 65)s and(1. 01 ± 0. 78)s, (100.58 ± 36. 56)s and (118. 65 ± 143. 71)s, which showed no statistically significant difference (P>0. 05). The duration of nystagmus was more than 60 seconds. The intensity of nystagmus turning to normal and lesion side were(45.58 ± 28.71)°/s and (20.42 ± 16. 64)°/s. The intensity turning to normal side was greater than lesion side obviously. The difference was statistically significant (P<0. 05). Twenty-three patients withright HSC-Cup, and 13 patients with left HSC-Cup were taken in count. They were treated with above methods and return visit a week later. Twenty-eight patients (77. 77%) were cured, 36 patients (100. 00%) were improved. There were 4 patients recurrence during the follow-up. CONCLUSION The direction and duration time of induced nystagmus are available to diagnose the HSC-Cup. The lesion side may determined according to the intensity of induced nystagmus. Lying position avoiding normal side and oral-taken betahistine mesilate tablets is an effective treatment methods for HSC-Cup.
Benign Paroxysmal Positional Vertigo ; complications ; diagnosis ; Face ; Humans ; Nystagmus, Physiologic ; Retrospective Studies ; Semicircular Canals ; Treatment Outcome ; Vertigo

AIM:To detect and compare the longitudinal mitral annulus diastolic velocity and time interval changes by pulsed Doppler tissue imaging(DTI)in patients with angina pectoris(AP)and myocardial infarction(MI),and to explore the value of mitral annulus diastolic velocities and time intervals for evaluation of global left ventricular diastolic dysfunction.METHODS:Fifty patients with established coronary artery disease were divided into AP group(16 cases)and MI group(34 cases).Sixteen age-matched healthy individuals served as the control group.The septum,lateral,anterior and inferior walls of the mitral annulus were displayed,and selected for DTI spectrum sampling.Peak early and late diastolic velocities and their ratio,time to the onset and peak of the early diastolic wave,and regional isovolumic relaxation time were measured,and the average values of the four mitral annular sites were calculated and presented as Em,Am,Em/Am,QEm,TEm and IVRTm,respectively.RESULTS:Compared with the control group,Em and Em/Am were significantly lower in both the AP and the MI groups(P＜0.01).Em was even lower in the MI group than that in the AP group(P＜0.01).QEm,TEm and IVRTm were significantly longer in the AP and the MI groups than those in control group(P＜0.01 or P＜0.05).IVRTm was even longer in the MI group than that in AP group(P＜0.01).IVRTm had significantly negative correlation with Em(r=-0.64,P＜0.01).CONCLUSION:Em,Em/Am,QEm,TEm and IVRTm as measured by pulsed DTI may be promising indexes for quantitative assessment of global left ventricular diastolic dysfunction in patients with coronary artery disease.Em and IVRTm may indicate the severity of ischemic myocardial damage.

Objective To construct a naive human Fab fragment phage display library,from which the anti-gp96 antibodies may be panned by the gp96 purified from the tissue of urothelial carcinoma in the urinary bladder and provide a basis to new therapy for the malignant tumors.Methods Peripheral blood lymphocytes were isolated from 800 ml of blood,which was obtained from four healthy blood donors.The heavy chain Fd and light chain cDNA synthesized from the total RNA of lympbocytes were amplified by PCR with variable regions 5' and 3' primers of heavy and light chain, and the amplification products were ligated into the phagemid vector pComb3, then the ligated sample was transformed into competent E.coli XL1-Blue by electroperation.The transformed cells were infected with VCSM13 helper phage to yield recombinant phage antibody Fabs.The phagemids abstracted from amplified E.coli were cut with endonucleases such as Sac Ⅰ,Xba Ⅰ,Xho Ⅰ and Spe Ⅰ, and both the phage antibody Fabs and phage-raids abstracted from amplified E.coil were amplified by PCR to monitor the insertion of the genes of light chain or heavy chain Fd fragment.The gp96 purified from the urothelial carcinoma tissue of the bladder by affinity chromatog-raphy on eoncanavalin-A sephnrose and DEAE-sephnrose ion exchange chromatography were utilized as antigens to process three rounds of panning to the original Fab antibody library.Results The quantity of total RNA and cDNA were qualified.By combination of light chain and heavy chain genes, an antibody library containing 6.6×106 clones was obtained, and both the cutting of enzymes and PCR showed that there were the genes of light chain or heavy chain Fd fragment in the phagemids.The gp96 protein was obtained from urothelial carcinoma tissue in the urinary bladder.After having been panned by gp96, the original antibody library gained enrichment by 68 times.Conclusion Utilizing the technology of phage surface display, specific antibody can be gained from the human naive Fab phage display library,which can be used for immunological therapy for tumors.

Objective: To construct the eukaryotic expression vector pSurp-EGFP regulated by the survivin gene promoter and to detect the specific expression of the promoter in human laryngeal cancer Hep-2 cells by green fluorescent protein assay.Methods: Thesurvivin gene promoter was generated by polymerase chain reaction(PCR) and the CMV promoter of the pShuttle vector replaced by the survivin gene promoter to generate the plasmid pSurp.The three plasmids pShuttle,pSurp and pEGFP-C1 were respectively double-enzyme digested so as to produce the plasmids pCMV-EGFP and pSurp-EGFP carrying the CMV or survivin promoter.The purified pCMV-EGFP and pSurp-EGFP were transfected into Hep-2 cell and vascular endothelial cell ECV304 using liposome transfection reagent and the expressions of EGFP detected by the fluorescent microscope.Results: Thesurvivin gene promoter was successfully cloned by PCR,and thesurvivin gene promoter-regulated pSurp-EGFP was constructed.Green fluorescence was observed in Hep-2 cells but not in ECV304. Conclusion: The high specific activity of the survivin gene promoter in Hep-2 cells that we successfully constructed attributes to the studies of tumor specific gene therapy.

Objective Genetic diversity and affinity relationships among 13 species of Dendrobium Sw.were analyzed,the result laid a solid foundation for the better use of this resources.Methods Random amplified polymorphic DNA(RAPD) technique was used to analyze genetic diversity,and the dendrogram was constructed by UPGMA.Results Ten RAPD primers were applied to do random amplification.A total of 188 DNA bands was detected,180 among which were polymorphic,the average rate of polymorphic bands was 95.74%.The result of cluster analysis by using UPGMA method showed that 13 genotypes could be classified into three types in genetic distance 0.63.This outcome was corresponding to the result by using traditional classification.Conclusion It is concluded that RAPD markers can be used on the studies of genetic relationships and classification of species of Dendrobium Sw.sensitively.

Objective To establish early diagnosis,treatment and prevention of tuberculosis infection following kidney transplantation.Methods Eighteen post-operative tuberculosis infections were identified among 1024 kidney transplantations performed in Beijing Chaoyang Hospital between January 2002 and December 2009.Triple immunosuppressive therapy strategy was used for these 18 patients.Of the 14 patients who received immune induction therapy,4 were treated with monoclonal antibody,and the other 10 were treated with anti-thymocyte globulin(ATG)or anti-lymphocyte globulin(ALG).Results The interval between renal transplantation and identification of tuberculosis infection ranged from 1 to 54 months.Posttransplant tuberculosis infection showed no typical clinical manifestations at early stage.Persistent or intermittent fever was the main symptom.High resolution CT and bronchoalveolar lavage fluid(BALF)test were useful tools for confirmed diagnosis.After routine anti-tuberculosis treatment,17 patients were cured and 1 patient died.Conclusions The early stage symptoms of post-transplant tuberculosis may be atypical,which could result in misdiagnosis.Pulmonary high-resolution CT examination and BALF test could provide strong evidence for tuberculosis infection.