Use of a novel induced spawning technique for the first reported captive spawning of Tetraodon nigroviridis

The spotted green pufferfish (Tetraodon nigroviridis) is an important genetics model animal due to its small, well-mapped genome. However, only wild-caught juveniles and adults are available to researchers. A lack of gametes, fertilized eggs, developing embryos, and other early life stages hampers development of the full potential of T. nigroviridis as a model research species. We report on successful spawning trials using a novel induced spawning technique, ovarian lavage. Chorulon^® (human chorionic gonadotropin, hCG) was injected into a catheter inserted into the oviduct at a rate of 3 μl/g body weight. In one trial, a female paired with a male spawned in an aquarium at about 72 h post-treatment. In other trials, females were hand-stripped of eggs at 36 h post-treatment. There were 3680 eggs/g of eggs and females produced up to 24% of their body weight in eggs. Hatch resulted from all trials on the 4th day post-fertilization. Ovarian lavage is a simple method for administering spawning hormones, uses a catheter technique similar to that frequently performed to determine egg maturity in broodstock, and eliminates the need for injection.     

炎症因子在大叶性肺炎患儿支气管肺泡灌洗液及血清中的表达及临床意义

目的:探讨炎症因子在大叶性肺炎患儿支气管肺泡灌洗液及血清中的表达水平及其临床意义。方法:收集2014年6月至2016年6月我院儿科收治的79例大叶性肺炎患儿为病例组,按照临床肺部感染评分(CPIS)分为重度组(n=46)和轻度组(n=33),于同期随机选取30例健康儿童为对照组,采用双抗体夹心酶联免疫吸附法(ELISA)检测各组支气管肺泡灌洗液及血清炎症因子如白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-17(IL-17)、肿瘤坏死因子(TNF-α)以及高迁移率族蛋白B1(HMGB1)表达水平。结果:重度组、轻度组血清IL-6、IL-8、IL-17、TNF-α及HMGB1水平高于对照组,重度组血清IL-6、IL-8、IL-17、TNF-α及HMGB1水平高于轻度组,差异有统计学意义(P0.05)。病例组患儿支气管肺泡灌洗液IL-6、IL-8、IL-17、TNF-α及HMGB1水平高于血清,差异有统计学意义(P0.05)。重度组、轻度组支气管肺泡灌洗液IL-6、IL-8、IL-17、TNF-α及HMGB1水平高于血清,差异有统计学意义(P0.05),重度组支气管肺泡灌洗液、血清IL-6、IL-8、IL-17、TNF-α及HMGB1水平分别高于轻度组,差异有统计学意义(P0.05)。经Spearman积矩相关分析,支气管肺泡灌洗液与血清中IL-6、IL-8、IL-17、TNF-α及HMGB1水平分别呈正相关关系(r=0.711、0.695、0.752、0.793、0.728,P0.05)。结论:炎症因子在大叶性肺炎患儿支气管肺泡灌洗液及血清中呈高表达,并与大叶性肺炎病情严重程度密切相关,联合检测有助于早期评估病情。     

Effect on fertility of human chorionic gonadotrophin and uterine lavage with oxytocin performed after mating in Arabian barren mares

The objective of the present study was to evaluate the beneficial effect of hCG injected immediately after mating in Arabian barren mares treated with uterine lavage and oxytocin. Arabian barren mares (n = 36) with PMIE were subjected to detailed clinical examinations including palpation per rectum, vaginoscopy, and cytological examination. After mating the 36 mares were randomly divided into four groups. The mares in group 1 (n = 10) were immediately after breeding injected with hCG 3000 IU IM. Uterine lavage with 1 L of N-saline containing 4 million IU of crystalline penicillin and 4 g of streptomycin sulphate was performed 4 h after breeding. Then mares received two injections of oxytocin 40 IU IM 2 h apart after 6 h of mating. Mares in group 2 (n = 10) treated with uterine lavage and oxytocin as group 1. While mares in group 3 (n = 10) received uterine lavage only. A control group (n = 6) as group 4 did not received any treatment. The results of clinical examination indicated that 69.4% of PMIE mares were harboring severe endometritis and 30.6% with a moderate form of endometritis. Significant (P < 0.01) increase in lymphocytes were founded in barren mares included in this study. Higher pregnancy rate (P < 0.01) was founded in Arabian barren mares 80% injected with hCG immediately after breeding and uterine lavage and oxytocin. No significant difference was found in mares received uterine lavage and oxytocin and uterine lavage only. In a conclusion, administration of hCG immediately after mating and intrauterine lavage containing antibiotics performed 4 h and two injections of oxytocin 40 IU IM 2 h apart after 6 h of mating had improved fertility of Arabian barren mares.     

The time course of responses to intratracheally instilled toxic Stachybotrys chartarum spores in rats

Stachybotrys chartarum is a fungal species that can produce mycotoxins, specifically trichothecenes. Exposures in the indoor environment have reportedly induced neurogenic symptoms in adults and hemosiderosis in infants. However, little evidence has linked measured exposures to any fungal agent with any health outcome. We present here a study that focuses on quantitatively assessing the health risks from fungal toxin exposure. Male, 10 week old Charles River-Dawley rats were intratracheally instilled with approximately 9.6 million Stachybotrys chartarum spores in a saline suspension. The lungs were lavaged 0 h (i.e., immediately post-instillation), 6, 24 or 72 h after instillation. Biochemical indicators (albumin, myeloperoxidase, lactic dehydrogenase, hemoglobin) and leukocyte differentials in the bronchoalveolar lavage fluid and weight change were measured. We have demonstrated that a single, acute pulmonary exposure to a large quantity of Stachybotrys chartarum spores by intratracheal instillation causes severe injury detectable by bronchoalveolar lavage. The primary effect appears to be cytotoxicity and inflammation with hemorrhage. There is a measurable effect as early as 6 h after instillation, which may be attributable to mycotoxins in the fungal spores. The time course of responses supports early release of some toxins, with the most severe effects occurring between 6 and 24 h following exposure. By 72 h, recovery has begun, although macrophage concentrations remained elevated.This revised version was published online in October 2005 with corrections to the Cover Date.     

Increased Hematopoietic Extracellular RNAs and Vesicles in the Lung during Allergic Airway Responses

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Acute exacerbations of COPD are associated with significant activation of matrix metalloproteinase 9 irrespectively of airway obstruction,emphysema and infection

Background

Acute exacerbations of chronic obstructive pulmonary disease (AE-COPD) are associated with accelerated aggravation of clinical symptoms and deterioration of pulmonary function. The mechanisms by which exacerbations may contribute to airway remodeling and declined lung function are poorly understood. In this study, we investigated if AE-COPD are associated with differential expression of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in bronchoalveolar lavage (BAL).

Methods

COPD patients undergoing diagnostic bronchoscopy, with either stable disease (n = 53) or AE-COPD (n = 44), matched for their demographics and lung function parameters were included in this study. Protein levels of MMP-2,–9,–12 and of TIMP-1 and -2 in BAL were measured by ELISA. Enzymatic activity of MMP-2 and -9 was assessed by gelatin zymography.

Results

We observed that MMP-9, TIMP-1 and TIMP-2 were significantly increased in BAL during AE-COPD. Furthermore, there was a significant negative correlation of MMP-9, TIMP-1 and TIMP-2 with FEV1% predicted and a significant positive correlation of TIMP-1 and TIMP-2 with RV% predicted in AE-COPD. None of MMPs and TIMPs correlated with DLCO% predicted, indicating that they are associated with airway remodeling leading to obstruction rather than emphysema. In AE-COPD the gelatinolytic activity of MMP-2 was increased and furthermore, MMP-9 activation was significantly up-regulated irrespective of lung function, bacterial or viral infections and smoking.

Conclusions

The results of this study indicate that during AE-COPD increased expression of TIMP-1, TIMP-2, and MMP-9 and activation of MMP-9 may be persistent aggravating factors associated with airway remodeling and obstruction, suggesting a pathway connecting frequent exacerbations to lung function decline.

Electronic supplementary material

The online version of this article (doi:10.1186/s12931-015-0240-4) contains supplementary material, which is available to authorized users.     

Cysteinyl-leukotriene 1 receptor antagonist attenuates bleomycin-induced pulmonary fibrosis in mice

Leukotrienes are lipid mediators of inflammation derived from the 5-lipoxygenase pathway of arachidonic acid metabolism, and recent evidence suggests that they play an important role in pulmonary fibrosis. Montelukast is a cysteinyl-leukotriene 1 receptor antagonist that has been found to reduce airway remodeling, including subepithelial fibrosis, in a murine model of asthma, but the therapeutic effect of montelukast on pulmonary fibrosis remains unclear. In this study, we investigated whether montelukast is capable of preventing bleomycin-induced pulmonary fibrosis in mice. On day 1, C57BL/6 mice were given a single intratracheal injection of bleomycin (2.5 mg/kg), and montelukast (1.0 mg/kg) or vehicle alone subcutaneously 2 h later and on days 1-5 of each week for two weeks. The total number of cells in bronchoalveolar lavage fluid (BALF) was reduced in the montelukast group on day 7 and on day 14, and cellular inflammation and fibrosis were attenuated on day 14 as indicated by significant decrease in the Ashcroft score and lung hydroxyproline content. Although cysteinyl-leukotriene level in BALF was not significantly different, transforming growth factor beta (TGFbeta) level in BALF by ELISA and TGFbeta expression in lung tissue by immunohistochemistry was reduced on day 14 in the montelukast group. The results of this study show that montelukast inhibits the inflammatory process and development of bleomycin-induced pulmonary fibrosis in mice and that these effects may be associated with a decrease in TGFbeta expression. They also suggest that montelukast may serve as a new therapy for patients with interstitial pulmonary fibrosis.     

Chemical modification-mediated optimisation of bronchodilatory activity of mepenzolate,a muscarinic receptor antagonist with anti-inflammatory activity

The treatment for patients with chronic obstructive pulmonary disease (COPD) usually involves a combination of anti-inflammatory and bronchodilatory drugs. We recently found that mepenzolate bromide (1) and its derivative, 3-(2-hydroxy-2, 2-diphenylacetoxy)-1-(3-phenoxypropyl)-1-azoniabicyclo[2.2.2]octane bromide (5), have both anti-inflammatory and bronchodilatory activities. We chemically modified 5 with a view to obtain derivatives with both anti-inflammatory and longer-lasting bronchodilatory activities. Among the synthesized compounds, (R)-(–)-12 ((R)-3-(2-hydroxy-2,2-diphenylacetoxy)-1-(3-phenylpropyl)-1-azoniabicyclo[2.2.2]octane bromide) showed the highest affinity in vitro for the human muscarinic M3 receptor (hM₃R). Compared to 1 and 5, (R)-(–)-12 exhibited longer-lasting bronchodilatory activity and equivalent anti-inflammatory effect in mice. The long-term intratracheal administration of (R)-(–)-12 suppressed porcine pancreatic elastase-induced pulmonary emphysema in mice, whereas the same procedure with a long-acting muscarinic antagonist used clinically (tiotropium bromide) did not. These results suggest that (R)-(–)-12 might be therapeutically beneficial for use with COPD patients given the improved effects seen against both inflammatory pulmonary emphysema and airflow limitation in this animal model.     

支气管肺泡灌洗术用于重症支气管哮喘的疗效评价及对肺功能、炎症因子水平的影响

目的:探讨支气管肺泡灌洗术用于重症支气管哮喘的疗效及对肺功能、炎症因子水平的影响。方法:选择我院2015年3月～2018年3月收治的150例重症支气管哮喘患者,均接受支气管肺泡灌洗术治疗,分析其临床疗效,临床症状改善状况,治疗前后肺功能、炎症因子、血气指标的变化情况和不良反应的发生情况。结果:治疗后,150例患者中,临床控制70例,好转60例,无效20例,总有效率为86.67%(130/150),哮鸣音消失时间为(5.17±0.64)d、ACT评分(27.85±3.86)分、住院时间为(5.73±0.75)d。治疗后,患者用力肺活量(FVC)、1s用力呼气容积(FEV1)、呼气峰流速(PEF)、血氧分压(PaO_2)、血氧饱和度(SaO_2)均显著高于治疗前(P0.05),嗜酸性粒细胞(EOS)、干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)及二氧化碳分压(PaCO_2)低于治疗前(P0.05)。150例患者中,发生支气管痉挛2例,咽喉部疼痛3例,无猝死、大咯血、气胸等严重并发症发生。结论:支气管肺泡灌洗术用于重症支气管哮喘的疗效肯定,可有效改善患者肺功能,并减轻炎症反应。     

Purification and characterization of a silica-induced bronchoalveolar lavage protein with fibroblast growth-promoting activity

Experimentally induced silicosis provides a good model for chronic interstitial pulmonary inflammation and fibrosis. In the present study, a specific single polypeptide with an apparent molecular mass of 58,000 and a pI of 4.5 was purified and characterized from the bronchoalveolar lavage fluid of silicotic rats. The same protein was also isolated from both the extract and conditioned medium of alveolar macrophages of silicotic rats. Therefore, this protein was termed an inducible silicotic (rat) bronchoalveolar lavage protein-p⁵⁸ (iSBLP⁵⁸) or an inducible silicotic (rat) pulmonary macrophage factor (iSPMF-p⁵⁸). iSBLP⁵⁸ has been purified to homogeneity by a combination of gel permeation, Mono Q ion exchange, and reverse-phase high performance liquid chromatography. This polypeptide displayed a potent fibroblast growth-promoting activity in vitro. The sequence of the first 15 NH₂-terminal amino acids was determined and was found to have high sequence homology with members of the mammalian chitinase-like protein family, which includes human cartilage gp39, mammalian oviduct-specific glycoprotein, and a secretory protein from activated mouse macrophages. J. Cell. Biochem. 67:257–264, 1997. © 1997 Wiley-Liss, Inc.