Objective:To obtain the functional single chain Fv antibody(scFv) against acidic isoferritin(AIF).Methods:An expression vector pPOW4c9 was constructed by subcloning AIF4c9 scFv gene into a heat-inducible bacterial expression plasmid pPOW3. Then recombinant vector was introduced into E.coli DH5? by electro-transformation. The soluble expression was performed by temperature induction. After purified by the Ni-chelating chromatography, the recombinant anti-AIF scFv was characterized.Results:Soluble expression of the scFv in E. coli was achieved. The yield of purified anti-AIF scFv was 1.6 mg/L. The recombinant protein recognized AIF specifically identified by ELISA and western blotting, and an affinity constant of scFv was 3.18?10~ -8 mol/L.Conclusion:The results indicate that recombinant soluble scFv retains the specific binding activity to AIF.

Curriculum ; Education, Medical, Graduate ; Education, Medical, Undergraduate ; Medicine, Chinese Traditional

To perform a systemic review and meta-analysis of the diagnostic accuracy of PET (CT) and metaiodobenzylguanidine (MIBG) for diagnosing neuroblastoma (NB),electronic databases were searched as well as relevant references and conference proceedings.The diagnostic accuracy of MIBG and PET (CT) was calculated for NB,primary NB,and relapse/metastasis of NB based on their sensitivity,specificity,and area under the summary receiver operating characteristic curve (AUSROC) in terms of per-lesion and per-patient data.A total of 40 eligible studies comprising 1134 patients with 939 NB lesions were considered for the meta-analysis.For the staging of NB,the per-lesion AUSROC value of MIBG was lower than that of PET (CT) [0.8064±0.0414 vs.0.9366±0.0166 (P＜0.05)].The per-patient AUSROC value of MIBG and PET (CT) for the diagnosis of NB was 0.8771±0.0230 and 0.6851±0.2111,respectively.The summary sensitivity for MIBG and PET (CT) was 0.79 and 0.89,respectively.The summary specificity for MIBG and PET (CT) was 0.84 and 0.71,respectively.PET (CT) showed higher per-lesion accuracy than MIBG and might be the preferred modality for the staging of NB.On the other hand,MIBG has a comparable diagnosing performance with PET (CT) in per-patient analysis but shows a better specificity.

OBJECTIVETo explore the clinical and epidemiological characteristics of serious cases of hand-foot-and-mouth disease (HFMD) infected by EV71, in order to provide scientific evidence for prevention and control of HFMD.

METHODSInformation was collected by questionaires through consulting medical cases. Data was input by Epidata, and analysed by software SAS 9.13.

RESULT201 severe cases were investigated. 84.65% of the cases were below 3 years old. The youngest one was 5 months and the oldest one was 8 years old. The ratio for male and female was 2.2: 1.85. 08% of the cases were distributed sporadically. 51.74% of them lived in rural, 29.36% of them lived in urban and 19.9% of them lived at the fringe area of rural and urban. 81.59% of the cases became serious between 1 and 4 days after infected. 100% cases had fever and 99.95% of them had a rash. 96.52% of them had nerve system symptoms. The main complications were virulent spinal encephalitis, pneumonia and breathing exhaustion. 98.01% of the patients were recovered or cured.

CONCLUSIONThe cases aged below 3 years old are high risk persons. Rural area and the fringe area of rural and urban are the key area for disease control. 1-4 days after onset is the key period to prevent complications.

Child ; Child, Preschool ; China ; epidemiology ; Enterovirus A, Human ; isolation & purification ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; pathology ; virology ; Humans ; Infant ; Male

COP9 Signalosome Complex ; Carcinoma, Hepatocellular ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; chemistry ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Gene Expression Regulation, Neoplastic ; Humans ; Intracellular Signaling Peptides and Proteins ; chemistry ; genetics ; metabolism ; Peptide Hydrolases ; chemistry ; genetics ; metabolism ; Phosphorylation ; Phosphoserine ; metabolism

0.05)between the two groups.Conclu- sion The long-term outcomes of e-CHB is not markedly different compared with that of e+CHB.

Objective To explore the expression of silencer of death domains(SODD) and its clinical significance and relationship with phospho-NF-?B-p65 proteins in bone marrow cells of acute lymphoblastic leukemia(ALL)in children,and the expression of SODD and phospho-NF-?B-p65 in Jurkat cells treated with chemotherapeutic drugs in order to find a new chemotherapeutic target.Methods The expressions of SODD and phospho-NF-?B-p65 proteins in bone marrow cells were detected by immunohistochemistry in 25 children with ALL.The apoptosis incidence was measured by Annexin-V-Fluorescence/PI double-labeling flow cytometry and the expression of SODD and phospho-NF-?B-p65 proteins were determined by Western blotting in Jurkat cells.Results It was found that the expression of SODD and active p65 expression in ALL were significantly higher than those in healthy control group.The expression of SODD and phospho-NF-?B-p65 proteins in the high-risk(HR) group was significantly higher than those in standard-risk(SR) group(Pa

OBJECTIVETo compare the angiogenesis in unstable and stable plaques and to investigate the potential role of neovessels in creating vulnerable sites for atherosclerotic plaques.

METHODSSpecimens of coronary arteries were obtained from 52 autopsy cases with acute coronary syndromes. Plaque morphology was studied by use of stained slides. 922 tissue blocks of late-stage lesions were classified into two groups: (1) unstable plaque (n = 153), the plaque was characterized by a large extracellular lipid core (more than 40% of the plaque area); (2) stable plaque (n = 769), lipid core less than 40% of the plaque area. Forty blocks were selected randomly from each group and serial sections were stained immunohistochemically with a polyclonal antibody against F VIII RAg. Computer-aided planimeter was used for quantitative analysis.

RESULTSIn unstable plaques, the occurrence of neovessels was more frequent and the neovessel density (number/mm(2)) was significantly increased as compared to that of stable plaques (frequency: 80.4% vs 66.6%, P < 0.01; shoulder: 22.16 +/- 19.96 vs 10.04 +/- 11.52, base: 21.68 +/- 20.44 vs 9.68 +/- 11.52, fibrous cap: 3.80 +/- 5.32 vs 1.48 +/- 2.28, P < 0.05). Most neovessels were located in the shoulder region and at the base of plaques.

CONCLUSIONSThese findings suggest that neovessels in coronary atherosclerotic plaques are closely associated with the decreased stabilization of the plaques.

Aged ; Aged, 80 and over ; Coronary Artery Disease ; pathology ; Coronary Vessels ; pathology ; Female ; Humans ; Male ; Neovascularization, Pathologic ; pathology

OBJECTIVETo evaluate the usefulness of new microspincolumn method for the measurement of a1pha-fetoprotein variant AFP-L3 in differentiation of benign and malignant liver disease and the warming for liver cancer.

METHODSAFP-L3 was isolated by using microspincolumn coupled with lens culinaris agglutinin (LCA), AFP and AFP-L3 were determined with chemiluminescent immunoassay, the proportion of AFP-L3 levels AFP-L3(%) were calculated, and the relationship between the elevated AFP-L3(%) levels and benign and malignant liver disease was analyzed.

RESULTSThe levels of AFP-L3(%) in serum of patients with hepatocellular carcinoma was significantly higher than those in the patients with other liver diseases (P < 0.001). Taking AFP-L3(%) >or= 10% as the diagnostic criteria, the sensitivity for diagnosis of liver cancer was 90.9%.

CONCLUSIONDetection of AFP-L3 seemed to be of clinical value in diagnosis and differential diagnosis of hepatocellular carcinoma; it may be especially important for identifying patients with hepatocellular carcinoma whose a1pha-fetoprotein level is low.

Adult ; Aged ; Carcinoma, Hepatocellular ; blood ; diagnosis ; Diagnosis, Differential ; Female ; Hepatitis, Chronic ; blood ; diagnosis ; Humans ; Immunoassay ; methods ; Liver Cirrhosis ; blood ; diagnosis ; Liver Neoplasms ; blood ; diagnosis ; Luminescent Measurements ; methods ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult ; alpha-Fetoproteins ; analysis

OBJECTIVESTo analyze the relationship between oxidized low density lipoprotein (oxLDL), angiogenesis and stabilization of atherosclerotic plaques in human coronary arteries; and to investigate the role of oxLDL in creating vulnerable sites in atherosclerotic plaques.

METHODSSamples of coronary arteries were obtained at autopsies of 42 patients with acute coronary syndrome. Eighty randomly selected blocks were studied by immunohistochemistry using antibodies against oxLDL and endothelial cells (factor VIII). Computer-aided planimeter was used for quantitative analysis.

RESULTSIn unstable plaques, percentage of immunoreactive areas for oxLDL was significantly higher than that in stable plaques. Most of the oxLDL were located in shoulder region of these plaques, as compared to the fibrous cap and basal regions. The details of distribution of oxLDL were as follows: shoulder region (20.43 +/- 3.12 for unstable plaques and 17.65 +/- 4.22 for stable plaques), fibrous cap (4.77 +/- 2.03 for unstable plaque and 2.80 +/- 0.22 for stable plaques) and basal region (5.65 +/- 1.65 for unstable plaques and 3.22 +/- 1.02 for unstable plaques). OxLDL was also a main component in the lipid core. In the shoulder region, there was a significant positive correlation between neovascularization and oxLDL (r = 0.8247, P = 0.000).

CONCLUSIONSThe amount of oxLDL is significantly higher in unstable atherosclerotic plaques, especially over the shoulder region. OxLDL in coronary atherosclerotic plaques is thus an important factor in determining stabilization of the plaques. OxLDL may induce influx of inflammatory cells which subsequently leads to decreased plaque stabilization.

Angina, Unstable ; metabolism ; pathology ; Coronary Artery Disease ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Lipoproteins, LDL ; metabolism ; Myocardial Infarction ; metabolism ; pathology ; Neovascularization, Pathologic ; metabolism ; pathology