Intranasal vaccination with ebola virus GP amino acids 258-601 protects mice against lethal challenge.

Ebola virus (EBOV) disease (EVD) leads to lethal hemorrhagic fever with a case fatality rate as high as 90%, thus posing a serious global public health concern. However, while several vaccines based on the EBOV glycoprotein have been confirmed to be effective in animal experiments, no licensed vaccines or effective treatments have been approved since the first outbreak was reported in 1976. In this study, we prepared the extracellular domain of the EBOV GP protein (designated as N20) by prokaryotic expression and purification via chromatography. Using CTA1-DD (designated as H45) as a mucosal adjuvant, we evaluated the immunogenicity of N20 by intranasal administration and the associated protective efficacy against mouse-adapted EBOV challenge in mice. We found that intranasal vaccination with H45-adjuvanted N20 could stimulate humoral immunity, as supported by GP-specific IgG titers; Th1 cellular immunity, based on IgG subclasses and IFN-γ/IL-4 secreting cells; and mucosal immunity, based on the presence of anti-EBOV IgA in vaginal lavages. We also confirmed that the vaccine could completely protect mice against a lethal mouse-adapted EBOV (MA-EBOV) challenge with few side effects (based on weight loss). In comparison, mice that received N20 or H45 alone succumbed to lethal MA-EBOV challenge. Therefore, mucosal vaccination with H45-adjuvanted N20 represents a potential vaccine candidate for the prevention of EBOV in an effective, safe, and convenient manner.

Development and characterization of a guinea pig model for Marburg virus.

The Angolan strain of Marburg virus (MARV/Ang) can cause lethal disease in humans with a case fatality rate of up to 90%, but infection of immunocompetent rodents do not result in any observable symptoms. Our previous work includes the development and characterization of a MARV/Ang variant that can cause lethal disease in mice (MARV/Ang-MA), with the aim of using this tool to screen for promising prophylactic and therapeutic candidates. An intermediate animal model is needed to confirm any findings from mice studies before testing in the gold-standard non-human primate (NHP) model. In this study, we serially passaged the clinical isolate of MARV/Ang in the livers and spleens of guinea pigs until a variant emerged that causes 100% lethality in guinea pigs (MARV/Ang-GA). Animals infected with MARV/Ang-GA showed signs of filovirus infection including lymphocytopenia, thrombocytopenia, and high viremia leading to spread to major organs, including the liver, spleen, lungs, and kidneys. The MARV/Ang-GA guinea pigs died between 7-9 days after infection, and the LD50 was calculated to be 1.1×10-1 TCID50 (median tissue culture infective dose). Mutations in MARV/Ang-GA were identified and compared to sequences of known rodent-adapted MARV/Ang variants, which may benefit future studies characterizing important host adaptation sites in the MARV/Ang viral genome.

[Safety and efficacy on cerebral protection with filter devices during carotid artery stenting in Chinese patients].

OBJECTIVE: To assess the efficacy, safety, procedural success and long-term clinical outcome in patients underwent percutaneous carotid stenting with distal device. METHODS: Percutaneous carotid stents with distal device were implanted to 58 patients with 59 significant (> 75%) carotid artery stenosis (49 men, mean age 68 years) between January 2000 to December 2005. Forty-five out of 58 patients were symptomatic, 35 had coronary artery diseases and 10 had previous strokes. RESULTS: Sixty one carotid stenting were implanted to 59 lesions in 58 patients. Stents with filter devices were successfully implanted in 57 out of 58 (98%) patients. Angioplasty success rate was 100%. Aspirin (300 mg/d) and Clopidogrel (75 mg x 2/d) were administered 3 days prior operation and clopidogrel was discontinued 30 days post stenting and aspirin was continued at dose of 100 mg/d. The percentage of stenotic carotid artery reduced from 85.3% to 6.2% after stenting and the diameter increased from 1.3 +/- 0.9 mm to 5.2 +/- 1.1 mm. Two minor strokes (3.4%) occurred during operation and at 14 days post stenting. All patients were discharged from the hospital after an average of 2.5 days hospitalization. At 14 +/- 2 months follow up, all patients survived and there were 2 asymptomatic restenosis (50% and 70% and the latter underwent successful balloon angioplasty), 2 myocardial infarctions (1 non-Q wave and 1 Q wave myocardial infarction, all underwent successful emergent PCI) and 2 minor strokes occurred at 6 and 8 months post stenting. CONCLUSION: Carotid stenting with distal device appears to be safe and effective in treating patients with carotid artery stenosis.

[Clinical analysis of 1,400 cases of coronary artery angiography without heparin].

OBJECTIVE: To evaluate the feasibility and safety of coronary artery angiography via femoral artery approach without heparin. METHODS: Totally 1 400 consecutive patients undergoing coronary artery angiography with heparin were analyzed retrospectively in comparison with the data of 354 patients for coronary artery angiography without heparin. RESULTS: Success in selective coronary artery angiography was achieved in 99.3% of the patients in the non-heparin group, with the mean operation time of 17.9+/-11.3 min ranging from 8 min to 1 hour. Angiography identified coronary artery stenosis in 72.2% of the patients including 49.8% with multivessel involvement. Subcutaneous hematoma occurred in 25 (1.8%) of the patients, and 1 (0.07%) patient developed arterial and venous fistula and pseudoaneurysm at the puncture site, but other complications (retroperitoneal hematoma, acute myocardial infarction, stroke and peripheral artery thrombotic events) occurred neither during nor after the procedure. In the heparin group, 18 (5.1%) patients developed subcutaneous hematoma and 1 (0.2%) had arterial and venous fistula and pseudoaneurysm at the puncture site after angiography. CONCLUSION: Coronary artery angiography without heparin is both safe and feasible with only very low risk of complications.

[Culture, purification and biological characters of olfactory ensheathing cells from adult transgenic mice].

Several studies have demonstrated the potential of olfactory ensheathing cells (OECs) for the repair of central and peripheral nerve injury. In this work, ensheathing cells that express the receptor gene coding for enhanced green fluorescent protein (eGFP) from adult mice were isolated and purified, and then, its biological characters were examined in vitro. The work was based on combinations of fluorescence confocal, phase contrast, cell proliferation assay and immunolabeling identification. The results showed that (1) two major morphologically and immunohistochemically distinct types of cells were present after 15 days in the primary cultures of adult transgenic mice olfactory nerves and glomerular layers of the olfactory bulb. One cell type was bipolar or multipolar OECs and strained positively for P75 low affinity neurotrophic receptor (P75N), S100, and glial fibrillary acidic protein(GFAP). The other type was fibroblasts with flat or endothelial-like shape, and reacted with antibody against Thy1.1. (2) A simple, inexpensive method for purifying ensheathing cells, in which various harvested cell types showed different rates of attachment to the uncoated culture ware, was developed. This technique neither binds any antibodies nor requires any costly equipment, and yields a large number of highly purified cells. (3) In sequential observations over 22 days in culture, the population of purified cells was maintained and continued to proliferate for longer. This experiment not only supported and advanced the ensheathing cell research but also offered ideal materials of in vivo transplantation for the repair of CNS injury.