Altered expression of 15-hydroxyprostaglandin dehydrogenase in chronic rhinosinusitis with nasal polyps / 临床耳鼻咽喉头颈外科杂志

Objective:To investigate the expression level and regulatory mechanism of 15-hydroxyprostaglandin dehydrogenase（HPGD） in chronic rhinosinusitis with nasal polyps（CRSwNP）. Methods:The expression pattern and level of HPGD in CRSwNP and control was observed using immunofluorescence, and western blot was used for analysis of HPGD expression in nasal polyp tissues. The effect of recombinant human high mobility group box-1（HMGB1） on HPGD expression in primary human nasal epithelial cells was observed, and the potential blocking effect of RAGE neutralizing antibody on HMGB1-induced HPGD expression was investigated. Results:The expression of HPGD was elevated in CRSwNP patients compared to the control, while the protein mainly localized at CD68-positive cells and epithelial cells. Recombinant human HMGB1 stimulated an increase in HPGD expression in primary human nasal mucosal epithelial cells at a time-dependent manner. Additionally, increased phosphorylation levels of MEK and elevated RAGE expression were also observed at 12 hours, but decreased at 24 hours after the incubation of HMGB1. The increase in the expression of HPGD induced by HMGB1 in primary human nasal epithelial cells was partly inhibited with RAGE neutralizing antibody. Conclusion:Elevated HPGD expression is observed in CRSwNP, predominantly in macrophages and epithelial cells. HMGB1 regulates HPGD expression through the RAGE-MEK signaling pathway, potentially providing a new target for future regulation of PGE2levels in CRSwNP.

Generation of dimeric single-chain antibodies neutralizing the cytolytic activity of vaginolysin

Background: Gardnerella vaginalis is a bacterial vaginosis (BV)-associated vaginal bacterium that produces the toxin vaginolysin (VLY). VLY is a pore-forming toxin that is suggested to be the main virulence factor of G. vaginalis. The high recurrence rate of BV and the emergence of antibiotic-resistant bacterial species demonstrate the need for the development of recombinant antibodies as novel therapeutic agents for disease treatment. Single-chain variable fragments (scFvs) generated against VLY exhibited reduced efficacy to neutralize VLY activity compared to the respective full-length antibodies. To improve the properties of scFvs, monospecific dimeric scFvs were generated by the genetic fusion of two anti-VLY scFv molecules connected by an alpha-helix-forming peptide linker. Results: N-terminal hexahistidine-tagged dimeric scFvs were constructed and produced in Escherichia coli and purified using metal chelate affinity chromatography. Inhibition of VLY-mediated human erythrocyte lysis by dimeric and monomeric scFvs was detected by in vitro hemolytic assay. The circulating half-life of purified scFvs in the blood plasma of mice was determined by ELISA. Dimeric anti-VLY scFvs showed higher neutralizing potency and extended circulating half-life than parental monomeric scFv. Conclusions: The protein obtained by the genetic fusion of two anti-VLY scFvs into a dimeric molecule exhibited improved properties in comparison with monomeric scFv. This new recombinant antibody might implement new possibilities for the prophylaxis and treatment of the diseases caused by the bacteria G. vaginalis.

Suplementação de vacas leiteiras com homeopatia: células somáticas do leite, cortisol e imunidade / Supplementation of dairy cows with homeopathy: milk somatic cells, cortisol and immunity

Avaliou-se o efeito da suplementação de uma combinação homeopática sobre a contagem de células somáticas do leite (CCS), o teor sanguíneo de cortisol e a resposta de anticorpos neutralizantes antivírus da raiva de vacas leiteiras. Trinta e duas vacas Holandesas em lactação foram blocadas em pares e aleatoriamente alocadas a um de dois tratamentos por 63 dias, posterior a um período de padronização de 14 dias. A CCS mensurada no final da padronização ajustou os valores semanais de CCS no modelo de análise estatística. Os tratamentos foram: 150 gramas de uma combinação homeopática (Hypothalamus, 10-30; Colibacilinum, 10-30; Streptococus Beta Hemolyticum, 10-60; Streptococus Uberis, 10-60; Phytolacca, 10-60; Calcium Phosphoricum, 10-30; Natrum Muriaticum, 10-60; Urtica Urens, 10-30; Silicea Terra, 10-400) em veículo mineral, ou 150 gramas do mesmo veículo mineral (controle). A homeopatia tendeu a aumentar a CCS de 124 para 222 x1.000 células mL-1 (P=0,09) e a CCS linearizada (P=0,08). Não foram detectados efeitos de tratamento sobre a concentração sérica de cortisol após estresse induzido por aspiração percutânea do saco ventral do rúmen (P=0,59) ou sobre o título de anticorpos neutralizantes em resposta à vacinação antivírus da raiva (P=0,40). A suplementação com homeopatia tendeu a aumentar a CCS de vacas com baixa CCS.

The effect of supplementing a homeopathic combination on milk somatic cell count (SCC), blood cortisol content and the antibody response to rabies vaccination of dairy cows was evaluated. Thirty-two lactating Holstein cows were paired blocked and randomly assigned to one of two treatments for 63 days, following a 14-day standardization period. The SCC measured at the end of standardization period adjusted weekly SCC values in the statistical analysis model. Treatments were: 150 grams of a homeopathic combination (Hypothalamus, 10-30; Colibacilinum, 10-30; Streptococcus Beta Hemolyticum, 10-60, Streptococcus Uberis, 10-60; Phytolacca, 10-60; Calcium Phosphoricum, 10-30; Natrum Muriaticum, 10-60; Urtica Urens, 10-30, Silicea Terra, 10-400) in mineral vehicle, or 150 grams of the same mineral vehicle (Control). Homeopathy tended to increase SCC from 124 to 222 x1,000 cells mL-1 (P=0.09) and linear SCC (P=0.08). There were no detectable treatment effects upon serum cortisol concentration following stress induced by percutaneous aspiration of the ventral rumen (P=0.59) and upon serum antibody title in response to rabies vaccination (P=0.40). The supplementation with homeopathy tented to increase the SCC of low SCC cows.