Objective To clone human chemokine ELC and express the ELC fusion protein. Methods Total RNA from human inflammatory tonsil was extracted and the cDNA was generated with reverse transcription. Mature ELC gene was amplified with PCR and NcoⅠand EcoRⅠ sites were added to the 5′ and 3′ terminal respectively, and then cloned into pET32a(+). E.coli DH5? was transformed with the recombinant plasmid, and positive clones were selected. The inserted DNA was verified by enzyme digestion and DNA sequencing. The fusion expression vector of mature ELC was formed with deletion mutation. ELC expression was analyzed by SDS-PAGE and Western blotting and the ELC fusion protein was purified. Results Human chemokine ELC was successfully cloned and the fusion protein was expressed and purified. Conclusion The ELC fusion protein was expressed with solubility.

BACKGROUNDThere are few reports discussing the surgical pathological characteristics of superficial endobronchial lung cancer (SELC) defined as cancer growth limited to the bronchial wall. Its prognosis and corresponding TNM staging have not been fully clarified. Little is known as to whether T status is impacted by the existence of associated atelectasis or pneumonia (which might be controversial, indicating either T1 or T2), and circumstantial invasion depth.

METHODSBetween 1988 and 2007, 81 out of 8817 surgically treated patients met SELC criteria; there was no detectable invasion beyond the bronchial wall. A retrospective review was performed and follow-up information was collected.

RESULTSThe overall five-year survival rate of 81 patients was 85.6%; for N0M0 (n = 67), N1M0 (n = 7) and N2M0 (n = 7) patients, they were 89.3%, 75.0% and 60.0%, respectively. Intraluminal tumor size measured from 0.4 to 3.0 cm; obstructive atelectasis or pneumonia was noted in 14 patients. The presence of tumor-associated obstructive atelectasis or pneumonia did not have a significant impact upon prognosis (P = 0.96), nor did the greatest diameter of the tumor (P = 0.70). Histology showed carcinoma in situ (level one) in 13 cases; invasion of the submucosal layer (level two) in 12, involvement of the muscular layer (level three) in 20, invasion into the space between the muscular layer and cartilage (level four) in 21, and bronchial cartilage infiltration in 15 (level five). In cases without lymphnode metastases, five-year survival was 100% for the first three levels and 84.0% and 61.3% for the level four and level five.

CONCLUSIONSRelative to TNM-based prognostic data, superficial endobronchial lung cancer exhibits increased five-year survival rates, and therefore should be placed at the forefront among tumors in the T1 class, regardless of tumor size or the presence of secondary obstructive atelectasis or pneumonia. Lymphnode metastasis is associated with a worse prognosis. Survival is negatively impacted by tumor infiltration depth into the bronchial wall.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; mortality ; pathology ; Female ; Humans ; Lung Neoplasms ; mortality ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Pneumonia ; mortality ; pathology ; Prognosis ; Pulmonary Atelectasis ; mortality ; pathology

OBJECTIVETo investigate the mechanism of enhanced large conductance calcium-activated potassium channel currents (BK) in coronary smooth muscle cells (SMCs) by docosahexaenoic acid (DHA).

METHODSCoronary SMCs were isolated by enzyme digestion. Potassium channels in coronary SMCs were identified by applications of different potassium blockers. Effects of DHA and its metabolite 16, 17-epoxydocosapentaenoic acid (16, 17-EDP) on BK channels in the absence and presence of cytochrome P450 epoxygenase inhibitor SKF525A were studied by patch clamp in whole-cell configuration.

RESULTSBK channels were widely distributed in SMCs, and BK currents in normal SMCs accounted for (64.2 ± 2.7)% of total potassium currents (n = 20). DHA could activate BK channels, and its 50% effective concentration (EC(50)) was (0.23 ± 0.03) µmol/L, however, the effect of DHA on BK channels was abolished after SMCs were incubated with cytochrome P450 epoxygenase inhibitor SKF525A. 16, 17-EDP, a metabolite of DHA, could reproduce the effects of DHA on BK channels, and its EC(50) was (19.7 ± 2.8) nmol/L.

CONCLUSIONDHA and metabolites can activate BK channels and dilate coronary arteries through activating cytochrome P450 epoxygenase pathway.

Animals ; Coronary Vessels ; cytology ; drug effects ; metabolism ; Cytochrome P-450 Enzyme Inhibitors ; Docosahexaenoic Acids ; pharmacology ; Fatty Acids, Unsaturated ; pharmacology ; Large-Conductance Calcium-Activated Potassium Channels ; metabolism ; Muscle, Smooth, Vascular ; drug effects ; metabolism ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Proadifen ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate two commercial anti-hepatitis E virus (HEV) IgM kits used for differential diagnosis of acute enteric viral hepatitis.

METHODSThe kit for IgM capture assay, was produced with a recombinant HEV structural protein protecting primates against experimental infection by different HEV genotypes, while the other kit for indirect ELISA was produced with recombinant structural proteins from different HEV genotypes. The serum specimens were taken from 241 cases with a confirmed or presumptive diagnosis of hepatitis A and 74 cases with a confirmed or presumptive diagnosis of hepatitis E.

RESULTSThe sensitivity and specificity of the IgM capture assay kit were 97% and 100%, respectively, and the corresponding values for the other kit were 70% and 78%, respectively.

CONCLUSIONThe IgM capture assay kit has higher sensitivity and specificity in diagnosing acute enteric viral hepatitis E.

Diagnosis, Differential ; Hepatitis E ; diagnosis ; immunology ; Humans ; Immunoglobulin M ; blood ; immunology ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

OBJECTIVETo evaluate the serological markers and biological marker in the diagnosis of hepatitis E infection in a rhesus monkey model.

METHODS86 rhesus monkeys had been infected with different doses of HEV. Hence, they were taken sequential blood samples at intervals up to 86 weeks for 4 hepatitis E virus (HEV) specific antibody assays (E2-IgM, E2-IgG, GL-IgG, and YES-IgG), and nucleic acid assay.

RESULTSAll the animals produced E2-IgG and all but one also produced E2-IgM and excreted the virus in stool, whereas positive rate of GL-IgG and YES IgG were low and correlated with virus level. Hepatitis occurred over a period of 4 weeks (between 3 an 7 weeks) after infection. Virological marker occurred mainly during incubation period and declined rapidly after onset of hepatitis. Seroconversion of E2-IgM occurred before onset of hepatitis in 70% monkeys and declined rapidly up to 50% of peak value after 4 weeks. E2-IgM seroconversion was closely paralleled by E2-IgG; however, E2-IgG persisted in all animals for the entire duration of experiment of up to 86 weeks. Production of GL-IgG and YES-IgG was delayed by one week after the E2 antibodies, these antibodies showed a transient occurrence and seroprevalence declined to 50% of the peak value over a period of 12 weeks.

CONCLUSIONE2-IgM might be used as a suitable acute hepatitis E marker, and E2-IgG as a suitable epidemiological marker. The seroconversion or titer elevation of GL-IgG and YES-IgG antibodies probably used to confirm the infection. The viral markers are optional for early diagnosis.

Alanine Transaminase ; blood ; Animals ; Biomarkers ; Genotype ; Hepatitis Antibodies ; blood ; Hepatitis E ; diagnosis ; Hepatitis E virus ; classification ; genetics ; immunology ; Immunoglobulin E ; blood ; Immunoglobulin M ; blood ; Macaca mulatta