血红素加氧酶-1介导CD4+CD25High调节性T淋巴细胞拮抗哮喘气道炎症的实验研究

探讨血红素加氧酶-1(HO-1, heme oxygenase-1)介导CD4⁺CD25^High调节性T淋巴细胞(Treg, regulatory T cells)拮抗哮喘气道炎症的作用. 用卵清蛋白(OVA)致敏、激发小鼠制备并建立哮喘动物模型, 并在致敏、激发过程中经氯化高铁血红素(Hemin)或锡-原卟啉(SnPP, Sn- protoporphyrin)处理. 分别测定激发后各组动物血清OVA特异性IgE, 支气管肺泡灌洗液中(BALF, bronchial alveolar lavage fluid)细胞总数和嗜酸性粒细胞(EOS, eosinophil)数及外周血CD4⁺CD25^High Treg细胞变化和肺组织HO-1和Foxp3 mRNA表达量, 结合病理切片分析气道炎症状况. 结果显示: OVA组、Hemin组、SnPP组血清OVA-特异性IgE和BALF中细胞总数及EOS数明显高于正常对照组, 但Hemin组IgE水平及BALF中细胞总数和EOS数明显低于OVA组; 而OVA组和SnPP组间IgE水平及BALF中细胞总数和EOS数无显著性差异; 病理组织学显示OVA组、Hemin组和SnPP组气道组织均见EOS浸润, 但Hemin组气道炎症仍明显轻于OVA组和SnPP组; Hemin组外周血CD4⁺CD25^High Treg细胞比例及肺组织Foxp3 mRNA相对表达量明显高于OVA组. Hemin显著上调HO-1表达. 实验表明, 用Hemin诱导HO-1高表达后外周血CD4⁺CD25^High Treg细胞比例及Foxp3 mRNA相对表达量明显升高, 同时血清OVA特异性IgE明显下降, BALF中细胞总数和EOS数减少, 气道炎症减轻; 提示HO-1可通过提高CD4⁺CD25^High Treg细胞比例并增强其功能来调节体内Th1/Th2平衡, 在支气管哮喘中起到保护作用.     

小青龙汤对小鼠支气管哮喘模型气道炎症及细胞因子的影响

目的:探讨中药方剂小青龙汤对小鼠哮喘模型气道炎症及细胞因子的影响。方法:40只BALB/c小鼠随机分为正常对照组(A组)、哮喘模型组(B组)、小青龙汤低剂量组(C组)和小青龙汤高剂量组(D组)。B、C、D组采用卵蛋白(OVA)腹腔注射致敏与雾化吸入激发制作哮喘模型。于OVA激发结束后24h收集支气管肺泡灌洗液(BALF)计数炎性细胞总数及嗜酸性粒细胞(EOS)数目,并测定BALF上清液中白细胞介素-4(IL-4)和干扰素-γ(IFN-γ)水平变化。结果:小青龙汤的干预治疗能显著降低小鼠BALF中炎性细胞总数及EOS数量;BALF上清液中IFN-γ水平明显升高,IL-4水平显著下降。D、C组与A组、B组有显著性差异(P<0.05),C组与D组结果亦存在显著性差异(P<0.05)。结论:小青龙汤能明显降低哮喘小鼠BALF中炎性细胞数量,影响细胞因子水平变化,从而改善哮喘气道炎症。     

小鼠哮喘发作过程中相关炎症因子的动态变化

目的:研究实验性哮喘小鼠模型在诱导哮喘发作不同时间点外周血中细胞因子IL-13、Eotaxin、MCP-1和TNF-α以及肺部浸润的炎症细胞数量变化。方法:将25只健康6-8周龄的雌性BALB/c小鼠随机分成模型组和对照组。利用卵清蛋白(OVA)诱导建立小鼠哮喘模型,模型组(Asthma)小鼠于第0、7天经腹腔注射卵清蛋白(OVA)致敏小鼠。第14-20天连续7天用1%OVA雾化激发小鼠哮喘发作,每次20 min,观察临床症状。正常对照组(Control)小鼠以0.9%NaCL代替0VA进行腹腔注射和雾化吸入。比较两组小鼠肺组织病理切片HE染色结果、肺泡灌洗液(BALF)中炎性细胞分类计数及细胞因子IL-13、Eotaxin、MCP-1和TNF-α的浓度变化。结果:模型组小鼠BALF中IL-13的水平在试验早期(致敏2天)即开始上升达68.9±4.34,此时Eotaxin和MCP-1未见明显升高;致敏7天时IL-13、Eotaxin和MCP-1均明显高于对照组,分别为88.3±3.39、67.4±4.24和38.9±3.1;激发1天组小鼠BALF中IL-13、Eotaxin和MCP-1浓度持续增高至最后一次激发后1天;而TNF-α在激发1天时出现明显升高达136.9±11.9,持续到最后一次激发后1天;从激发1天肺泡灌洗液染色显微镜下观察明以淋巴细胞和嗜酸性粒细胞浸润为主。肺组织HE染色显示哮喘组小鼠气道上皮有不同程度脱落,支气管平滑肌显著增厚,血管周围水肿,炎性细胞浸润。结论:在哮喘发生过程中,IL-13水平在致敏初期即开始升高,随着继续给予OVA,Eotaxin和MCP-1水平呈现显著增高;并伴随越来越多炎症细胞在肺部浸润,TNF-α水平出现缓慢增高,进而加重哮喘发作。     

补肾清肺方对小鼠支气管哮喘模型的影响

目的:探讨中药复方补肾清肺方对小鼠哮喘模型气道炎症及细胞因子的影响。方法:40只BALB/c小鼠随机分为正常对照组(A组)、哮喘模型组(B组)、补肾清肺方低剂量组(C组)和补肾清肺方高剂量组(D组)。B、C、D组采用卵蛋白(OVA)腹腔注射致敏与雾化吸入激发制作哮喘模型。于OVA激发结束后24h收集支气管肺泡灌洗液(BALF)计数炎性细胞总数及嗜酸性粒细胞(EOS)数目,并测定BALF上清液中白细胞介素-4(IL-4)和干扰素-γ(IFN-γ)水平变化。结果:补肾清肺方的干预治疗能显著降低小鼠BALF中炎性细胞总数及EOS数量;BALF上清液中IFN-γ水平明显升高,IL-4水平显著下降。D、C组与A组、B组有显著性差异(P<0.05),C组与D组结果亦存在显著性差异(P<0.05)。结论:补肾清肺方能明显降低哮喘小鼠BALF中炎性细胞数量,影响细胞因子水平变化,从而改善哮喘气道炎症。     

秀丽线虫天然粗提物对哮喘治疗效果研究

探讨秀丽隐杆线虫天然粗提物的提取及对哮喘动物治疗效果。将线虫经离心、洗涤、超声乳化、去除内毒素等方法获得线虫粗提物(CEC),以鸡卵清蛋白(OVA)致敏BALB/c雌性小鼠建立哮喘动物模型,用CEC与OVA混合物(CE)对哮喘动物治疗,以缓冲液(PBS)和OVA为对照,用ELISA法检测血清Ig E、Ig G1、Ig G2a抗体,分类计数支气管肺泡灌洗液(BALF)细胞,切取肺组织观察病理。研究显示,CE组与OVA组比较,血清Ig E、Ig G1、Ig G2a差异有统计学意义(P0.05),BALF中细胞总数、嗜酸性粒细胞、巨噬细胞等差异有统计学意义(P0.05)。线虫天然粗提物对哮喘动物有一定治疗效果。     

婴儿型双歧杆菌对哮喘小鼠气道炎症的作用研究

目的探讨婴儿型双歧杆菌对气道过敏小鼠的气道炎症作用及其机制。方法 6周龄BALB/c雄性小鼠40只,随机分为4组:阴性对照组、哮喘组、婴儿型双歧杆菌预防组和婴儿型双歧杆菌治疗组。哮喘组、预防组和治疗组用卵清蛋白(Ovalbvmin,OVA)进行致敏和激发诱发哮喘,阴性对照组用生理盐水进行致敏和激发。预防组第0至14天灌胃菌液;治疗组第15至28天灌胃菌液;阴性对照组和哮喘组全程灌胃生理盐水作对照。观察小鼠气道过敏表现;计数肺泡灌洗液(Bronchoalveolar lavage fluid,BALF)中细胞总数评估气道炎症程度;肺组织行HE染色;ELISA法测定血清中IL-10、OVA特异性IgE和OVA特异性IgG1的浓度以及肺泡灌洗液中细胞因子IL-4、IL-5、IL-10、IL-13和IFN-γ的浓度。结果 (1)哮喘组小鼠在激发后出现搔抓口鼻、竖毛、打喷嚏、腹肌扇动、弓背直立、二便失禁以及体重下降等表现,预防组和治疗组的症状较轻。(2)预防组和治疗组BALF细胞数量明显低于哮喘组(P0.05)。(3)肺组织HE染色后,哮喘组小鼠肺组织出现明显的炎症细胞浸润,预防组和治疗组肺组织炎症明显减轻。(4)预防组和治疗组血清OVA特异性IgE浓度明显低于哮喘组(P0.05),并且预防组低于治疗组(P0.05);治疗组血清OVA特异性IgG1浓度明显低于哮喘组和预防组(P0.05);预防组和治疗组血清IL-10浓度明显高于哮喘组(P0.05),且治疗组高于预防组(P0.05);(5)预防组和治疗组肺泡灌洗液中IL-4、IL-13含量明显低于哮喘组(P0.05);IL-5、IL-10、IFN-γ各组浓度都较低,未测出有效浓度。结论口服婴儿型双歧杆菌可以减轻过敏症状,降低肺组织炎症浸润程度,抑制Th2免疫反应。     

核心蛋白聚糖对慢性哮喘小鼠气道和肺血管胶原沉积的影响

目的:研究核心蛋白聚糖(decorin)对哮喘小鼠气道及肺血管胶原沉积的影响.方法:30只雌性昆明系小鼠随机分为对照组、慢性哮喘组及decorin干预组,每组10只.慢性哮喘组和decorin干预组小鼠第0、7、14d腹腔注射卵蛋白(OVA)致敏;第22d始雾化吸入2.5％OVA溶液激发哮喘,每次30min,每周3次,连续8周;decorin干预组小鼠每次OVA激发前2h腹腔注射decorin(0.25mg·kg-1)干预;对照组全部以生理盐水代替.末次激发24h后处死小鼠.酶联免疫吸附测定(ELISA)法测定小鼠血清和肺泡灌洗液(BALF)中转化生长因子-β1 (TGF-β1)水平;苏木精-伊红(HE)染色及Masson三色染色观察肺组织切片病理改变及胶原沉积;图像分析软件测定气道血管胶原沉积.结果:慢性哮喘组血清及BALF中TGF-β1水平显著高于对照组(P均＜0.01),而decorin 干预组较慢性哮喘组明显降低(P均＜0.01);慢性哮喘组气道壁胶原沉积厚度及肺血管壁胶原沉积厚度显著高于对照组(P均＜0.01),而decorin干预组上述改变较慢性哮喘组明显减轻(P均＜0.01).结论:Decorin干预可减轻哮喘小鼠气道及肺血管胶原沉积.     

不同浓度卵蛋白变应原对小鼠哮喘模型建立的影响

目的研究不同浓度卵蛋白(ovalbumin,OVA)变应原对小鼠的哮喘造模影响。方法 96只6～8周龄SPF级雌性BALB/c小鼠随机分为8组,分别为PBS组(对照组)、10μg组(A组)、20μg组(B组)、50μg组(C组)、100μg组(D组)、200μg组(E组)、500μg组(F组)、1000μg组(G组)。A～G组分别用含1%明矾的PBS配制相应浓度的OVA于第0、7和第14天对小鼠进行腹腔注射。于第21～27天连续7 d用含1%的OVA的PBS溶液雾化吸入激发各组小鼠。正常对照组使用PBS溶液致敏和激发。最后一次雾化吸入激发后24 h内,计数各组小鼠支气管肺泡灌洗液(BLAF)中嗜酸性粒细胞的含量,ELISA法检测IL-4、IL-5的分泌量及其血清IgG2a、IgE抗体的水平;肺组织病理切片观察各组小鼠哮喘模型的效果,评价最优哮喘造模的OVA浓度。结果 A～G组小鼠肺泡灌洗液中IL-4、IL-5含量均高于正常对照组(P<0.01),细胞因子水平随着OVA浓度的增高而逐渐下降;A～G组小鼠肺泡灌洗液中嗜酸性粒细胞数均高于正常对照组(P<0.01),从低浓度组至高浓度组嗜酸性粒细胞数从高向低变化;A～G组小鼠血清中总抗体IgE的水平均显著高于正常对照组(P<0.01),且随着OVA浓度的增高IgE水平逐渐下降。血清中IgG2a的水平则随OVA给药浓度的增高而逐渐增高;低浓度OVA致敏组小鼠肺组织标本可观察到明显的炎症浸润性病理表现,而高浓度组肺部组织病理变化不明显。结论低浓度的OVA连续致敏小鼠造成过敏性哮喘病理改变较为明显,随着OVA浓度的增高,造模效果逐渐降低,而高浓度的OVA则会导致模型小鼠发生免疫耐受。     

色胺酮对哮喘豚鼠体内NO和NOS水平的影响

目的:观察色胺酮对哮喘豚鼠血清及肺泡灌洗液中NO、NOS浓度的影响.方法:将初筛合格的48只豚鼠随机分成6组:空白组(A组)、慢性模型对照组(B组)、色胺酮高剂量组(C组)、色胺酮中剂量组(D组)、色胺酮低剂量组(E组)和阳性对照组(F组).采用磷酸卵蛋白(OVA)腹腔注射致敏加雾化吸入激发的方法复制豚鼠哮喘模型,造模成功后给予色胺酮和地塞米松治疗.测定各组豚鼠肺部组织、血清及肺泡灌液(BALF)中NO和NOS浓度并进行组间比较.结果:与B组相比,C、D、E、F组血清中NO和NOS浓度显著降低,组间比较,差异存在统计学意义(P<0.05);各组肺组织中NOS表达呈阴性;C、D、E、F组BALF中NO浓度较B组降低,组间比较,差异存在统计学意义(P<0.05).结论:色胺酮可降低哮喘豚鼠血清中NO和NOS水平,降低BALF中NO水平,可能对哮喘有一定的防治作用.     

口服柔嫩梭菌对OVA致敏小鼠气道炎症的影响

目的 在动物水平探索口服柔嫩梭菌(Clostridium leptum)对哮喘小鼠气道炎症的影响.方法 建立OVA致敏的BALB/c小鼠哮喘模型,依据检测气道炎症情况和气道反应性确定哮喘模型构建成功.30只BALB/c小鼠随机分为3组:正常对照组,安慰剂组和口服柔嫩梭菌治疗组,每组10只.通过HE染色检测小鼠肺组织病理变化,细胞计数检测肺泡灌洗液中炎性细胞(嗜酸性粒细胞、中性粒细胞、淋巴细胞、巨噬细胞)的数目,ELISA方法检测肺泡灌洗液中炎性因子(IL4、IL-5、IL-13)的表达情况,并检测各组小鼠的气道反应性.结果 验证OVA致敏哮喘小鼠模型构建成功;口服柔嫩梭菌可显著减轻OVA致敏小鼠的气道高反应性,气道炎症细胞浸润和炎性细胞因子分泌(P＜0.05).结论 口服柔嫩梭菌可显著减轻OVA致敏小鼠的气道炎症和气道高反应性,这可能为哮喘治疗提供新思路.     

Relative and combined effects of ethanol and protein deficiency on strontium and barium bone content and fecal and urinary excretion

To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.     

白蚁与动物及微生物的共生类型及其机制

综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生（宾主共栖和异种共栖）、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌（原生动物、细菌、真菌和放线菌）和外生菌（蚁巢伞菌等）间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用（如纤维素酶）及今后研究物种间的协同进化提供了基础资料。     

New amino-dithiaphospholanes and phosphoramidodithioites and their rhodium and iridium complexes

New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn₂NPSCH₂CH₂S^a(P-S^a) (Bn = benzyl, 4), Bn₂NPSCHCHS^a(CH₂)₃C^aH₂(P-S^a)(C^a-S^a) (6) and Bn₂NP(4-XC₆H₄OMe)₂ (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)₂] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl₂Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the ν_CO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The ¹J_PRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the ¹J_PSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh₃. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh₃.     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Rapporteur report: Basics and technology and metrology and standards

The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.     

Prostaglandin and thromboxane production by human and guinea-pig macrophages and leucocytes

The ability of partially purified human and guinea-pig haematogenous cell populations, when cultured in vitro, to metabolise arachidonic acid (AA) has been studied. Supernatants from 24 hour cell culture have been subjected to analysis for products of AA metabolism by gas chromatography with electron-capture detection.The cell types studied were human peripheral blood monocytes (both glass adherent and non-adherent), neutrophils, eosinophils and leukemic leucocytes; thoracic duct lymphocytes and lung alveolar macrophages. From the guinea-pig, induced and non-induced macrophage or neutrophil enriched peritoneal exudate populations, lymph node cells, peritoneal eosinophils and peripheral blood platelets were examined. Supernatants were assayed for the presence of PGE₂, PGD₂, PGF_2α, TXB₂ and 6-keto-PGF_1α. In all types studied PGE₂ and TXB₂ were the major products formed. The identification of PGE₂ and TXB₂ was confirmed by GC/MS with multiple ion monitoring.The results have been compared with other reports and their possible significance discussed in relation to the proposed role of prostaglandins as mediators and modulators in immunopathology.     

Cytoskeleton and mitochondrial morphology and function

It has been well established that the cytoskeleton is an essential modulator of cell morphology and motility, intracytoplasmic transport and mitosis, however cytoskeletal linkage to the organelles has not been unequivocally demonstrated. Indeed, cytoskeleton appears to be essential in determining and modulating gene phenotype as a function of cellular environment. According to recent studies, the organization of the cytoskeleton network together with associated protein(s) could be essential in regulating mitochondrial function and particularly the permeability of the mitochondrial outer membrane to ADP. The aim of this chapter is to summarize the main properties of the cytoskeletal environment of mitochondria and the possible role(s) of this network in mitochondrial function in myocytes.     

Cross-reactivity between storage and dust mites and between mites and shrimp

Many patients have sensitivities to multiple species of storage and house dust mites. It is not clear if this is because patients have multiple sensitivities to species-specific mite allergens or if these mites share many cross-reacting allergens. Our objective was to further define the cross-allergenicity between several species of storage and house dust mites using crossed-immunoelectrophoresis (CIE), crossed-radioimmunoelectrophoresis (CRIE), immunoblotting, and ELISA. CIE and CRIE reactions revealed that storage mites shared two cross-antigenic molecules and one of these bound IgE in a serum pool from mite allergic patients. Antibody in anti-sera built to each species of mite recognized many SDS–PAGE resolved proteins of other mite species and this suggested the potential for other cross-reactive allergens. Among patient sera, IgE bound to many different proteins but few had IgE that bound to a protein with common molecular weights across the mite species and this suggested mostly species-specific allergens. Antiserum built to each mite species precipitated one protein in shrimp extracts that bound anti-Der p 10 (tropomyosin) and IgE in the serum pool. Anti-Der p 10 showed strong binding to shrimp tropomyosin but very little to any of the mite proteins. ELISA showed the mite extracts contained very little tropomyosin. The storage and dust mites investigated contain mostly species-specific allergens and very small amounts of the pan-allergen tropomyosin compared to shrimp and snail.