Objective To establish a time resolved fluoroimmunoassay (TRFIA) method for detecting Glypican 3 (GPC3) and to explore the diagnostic value of serum GPC3 for hepatic carcinoma (HCC). Methods Microplate coated with anti-GPC3 monoclonal antibody 7C8 and GP9 labeled with Eu3+ were used to establish TRFIA kit. The serum concentrations of GPC3 in 41 HCC patients and 44 chronic hepatitis (CH) patients were quantitatively analyzed. AFP was detected by with lowest limit of 2.06 μg/L. The CV of inter and intra assay were 12.25% and 12.91%, respectively. The average serum concentration of GPC3 in HCC patients was (86.68±110.39) μg/L (median: 56.98 μg/L). But in CH patients it was only (14.77±29.48) μg/L, which was significantly lower than that in HCC (Wilcoxon W=1335.00, Z=-4.99, P<0.001). With diagnostic cut-off value set at 42.94 μg/L, the diagnostic sensitivity and specificity of TRFIA GPC3 for HCC were 58.5% (24/41) and 95.5%(42/44) respectively. The diagnostic sensitivity of AFP was 46.3% (19/41) in 41 HCC patients, and was raised to 78.0% (32/41) when combined with GPC3. Conclusions Serum GPC3 assay by TRFIA is established and it could increase the diagnostic sensitivity for HCC when combined with AFP.

Accommodation of the human eye ian extremely complex and dynamiprocess,which iaccomplished by the interaction between the central nervousystem and variouoculastructurethaare relevanto accommodation.Varioumechanismof accommodation have been puforward since the beginning of the 19th century,among which Helmhohz'theory ithe mosfamous.However,iistill challenged by othetheories.So far,the mechanism of accommodation hanobeen fully understood.The mosdirecmethod to study accommodation ito observe changein the biometry of the oculastructureduring accommodation,which ialso the mosobjective interpretation of accommodative mechanisms.The rapid developmenof imaging technologiein regardto ophthalmology makethipossible.Thiarticle aimto describe the use of variouimaging technologiein oculaaccommodative studiein vivo from the perspective of morphology.

Background Clinical studies indicated that the pathogenesis of most corneal dystrophy is associated with the mutation of the transforming growth factor beta-induced (TGFBI) gene.However,the molecular mechanism of mutated TGFBI gene in corneal dystrophy is unclear. Objective The present study was to investigate the expression of the TGFBI gene in human corneal tissue and cells in vitro.MethodsHuman corneal epithelial cells and keratocytes were cultured and passaged,and donor corneal tissue was obtained for the section preparation.RT-PCR was used to detect the expression of TGFBI mRNA in human corneal tissue and cells.Immunofluorescence was used to test the expression of the TGFBI protein in the human corneal tissue,and immunohistochemistry was used to test the expression of the TGFBI protein in human corneal epithelial cells and corneal stromal cells.ResultsRT-PCR analysis showed that TGFBI mRNA could be detected as a 1274 bp band in human corneal tissue and corneal stromal cells,but no TGFBI mRNA was observed in corneal epithelial cells.Immunofluorescence assay revealed that corneal stromal cells were positive ly expressed for the TGFBI protein,but the corneal epithelial cells did not express the TGFBI protein.Immunohistochemistry indicated that the expression of TGFBI was detected the red fluoressence in the cytoplasm of corneal stromal cells;however,no positive response was found in corneal epithelial cells.ConclusionsThe expression of the TGFBI gene occurs in human corneal stromal cells but not in the corneal epithelial cells.This result might be of helpful for studying the function and role of TGFBI gene in pathogenesis of corneal dystrophy.

Objective To analyse the feasibility of detecting F1 antibody to Yersinia pestis in flushing fluid of heart blood of Rhombomys opimus with enzyme linked immunosorbent assay(ELISA) method and its application value in surveillance of the disease. Methods Serum, flushing fluid of heart blood and infusion fluid of liver and spleen of Rhombomys opimus, which were caught by capture in the plague focus of Zunger basin in 2007, were taken to carry out detection for F1 antibodies to Yersinia pestis with ELISA method. The data were processed with SPSS 17.0. Results Positive rate and average titer of serum were 12.35%(11/162) and 25.35, of flushing fluid of heart blood were 10.49%(17/162) and 23.75 and of the infusion fluid of liver and spleen 6.79%(17/162) and 2240,respectively. No statistical difference was found in positive detection rate when it was compared between serum and flushing fluid of heart blood(χ2 = 1.333, P ＞ 0.05), but it was obviously different between serum and infusion fluid of liver and spleen(χ2 = 7.111, P ＜ 0.01 ) and between flushing fluid of heart blood and infusion fluid of liver and spleen(x2 = 6.250, P ＜ 0.05). There was a significant difference in average titer between serum, flushing fluid of heart blood and infusion fluid of liver and spleen(t = 2.290, 3.612, P ＜ 0.05 or ＜ 0.01 ). The plague F1 antibody positive coincidence rate of serum and flushing fluid of heart blood was 85.0%(17/20), of serum and infusion fluid of liver and spleen was 55.0% (11/20), and of flushing fluid of heart blood and infusion fluid of liver and spleen was 64.7%(11/17). Conclusions The ELISA method can detect Fl antibody in flushing fluid of heart blood,and the method is feasible in plague surveillance.

CCK correlates with the generation and progression of pancreatic cancer. The research aims to construct eukaryotic expression plasmid pIRES2-EGFP/CCK (CCK pDNA) and transiently express it in COS-7 cells. Total RNA was extracted from porcine intestinal mucosa. RT-PCR was used to amplify the aimed segments CCKcDNA which was then digested with EcoR1 and BamH1 and inserted into a eukaryotic expression plasmid pIRES2-EGFP to construct CCK pDNA. The constructed plasmid was transfected into COS-7 cells by lepofectamine TM2000-mediated transfer method.The expression of CCK in transfected COS-7 cells was detected 24, 48 and 72 h post-transfection with fluorescence microscopy and the expression level of CCK mRNA in transfected COS-7 cells was assayed by using RT-PCR. The results showed CCK pDNA was successfully constructed and expressed transiently in COS-7 cells. Green fluorescent protein could be detected in the COS-7 cells transfected with porcine CCK pDNA 24 h post-transfection. At 48th h post-transfection, the number of positive cells was increased significantly and much brighter green fluorescence could be detected.And 72 h post-transfection, the green fluorescence of positive cells became even stronger, while no green fluorescence was detected in the control group. The expression of CCK mRNA in the cells was detectable by using RT-PCR. In COS-7 cells transfected with CCK pDNA a high level of porcine CCK mRNA was detected while no expression of porcine CCKmRNA was found in the cells transfected with null plasmid. It was concluded CCK pDNA was expressed successfully in COS-7 cells,which lays a foundation for further research on the relationship between CCK and tumor.

Objective To compare the effect of three methods in diagnosis of plague by detecting of Yersina pestis F1 antigen. Methods In natural foci of plague, wild animal samples, such as blood, liver, spleen,and lymphoid tissue were collected, and the three methods of enzyme linked immunosorbent assay (ELISA),reverse indirect hemagglutination assay(RIHA) and gold-immunochromatography assay(GICA) were employed to detect F1 antigen of Yersina pestis. Results Total of 414 infused organ samples of natural death and captured wild animals in natural foci of plague were determined. Positive samples detected by GICA and ELISA were the same,the positive rates were 5.31%(22/414), both positive and negative coincidence rates were consistently 100%. Only 18 samples were positive by retrial in 186 samples with more than 2 holes aggregation by preliminary examination of RIHA, with nonspecific agglutination rate of 40.6% (168/414) and positive rate of 4.35% (18/414). The positive coincidence rate was 81.82% (18/22) between RIHA with GICA and ELISA, and negative coincidence rate was statistically significant(t = 4.379, P ＜ 0.01). Conclusions ELISA, RIHA and GICA can be used for early diagnosis of plague by detecting F1 antigen. The results of RIHA have quantitative significance, with higher non-specific agglutination rate, and heavy workload of re-examination; GICA and ELISA has the same specificity and sensitivity, but the results of GICA is only qualitative. ELISA excluded the defect of RIHA and GICA, and combines the advantages of both methods.

Objective To compare the diagnostic value of renal ultrasound scan (RUS) and 99Tcmdimercaptosuccinic acid (DMSA) renal scintigraphy in children with acute pyelonephritis (APN). Methods In all, 165 children with initial clinical diagnosis of APN, aged from 1.5 months to 11 yrs ( median 20 months), were included in the study, all of which were examined with RUS and DMSA renal scientigraphy. The diagnosis with DMSA renal scientigraphy results was taken as the standard reference to evaluate the diagnostic sensitivity and specificity of RUS. Results Of 99 out of all 330 kidneys that were found abnormal on DMSA renal scientigraphy, 31 were abnormal on RUS. Of the rest normal kidneys on DMSA scans renal scientigraphy, 4 were abnormal on RUS. Thus diagnostic sensitivity of RUS for APN was 31.3%(31/99) and specificity was 98.3% (227/231). Conclusions Although RUS provides with high diagnostic specificity for children with APN, its low sensitivity may underestimate the clinical evaluation of APN.More often than not, 99Tcm-DMSA renal scientigraphy is a clinical necesscity for the definite RUS diagnosis.