黄芪甲苷对变应性鼻炎小鼠线粒体动态平衡及NK 细胞分化影响的研究

目的：探讨黄芪甲苷对变应性鼻炎（AR）小鼠NK细胞分化的影响及机制。方法：C57/BL6小鼠随机分为空白组、AR模型组、黄芪甲苷组。观察各组小鼠行为学表现、鼻黏膜炎症反应;ELISA法测定血清IL-4、IL-5、IFN-γ水平;免疫荧光法检测各组NK细胞IL-4、IFN-γ表达;透射电镜等技术观察小鼠线粒体形态。结果：与AR模型组比较,黄芪甲苷组小鼠鼻过敏症状、鼻黏膜炎症明显改善,血清IL-4、IL-5水平降低。黄芪甲苷干预后,OVA致敏小鼠NKp46细胞IL-4、p-Drp1(Ser616)阳性比率降低。同时,黄芪甲苷组小鼠鼻黏膜NK细胞线粒体管状形态较AR模型组有所恢复,线粒体长度明显增加。结论：黄芪甲苷可能通过调节Drp1活性,恢复线粒体动态平衡,抑制NK细胞的NK2分化偏移,发挥改善AR的效应。     

γ-维生素E对变应性鼻炎大鼠炎症因子表达的影响

目的探讨γ-维生素E对变应性鼻炎大鼠(AR)Th1型和Th2型主要炎症因子的表达影响。方法 SD大鼠30只,随机分为γ-维生素E组、AR模型组和阴性对照组,每组10只。AR组和γ-维生素E采用卵白蛋白(OVA)联合A1(OH)_3致敏法建立AR大鼠模型,对照组采用生理盐水,γ-维生素E组于激发21d后,给予OVA的同时口服2.5mgγ-维生素E,连续7d;各组于给药后30min内观察各组大鼠临床表现,于最后一次激发后48h,取大鼠鼻腔分泌物,用ELISA试剂盒和Western-blot法检测IFN-γ、IL-17、IL-4和IL-5含量变化。结果 AR组大鼠均表现出打喷嚏、鼻痒和流鼻涕等AR的临床表现,而γ-维生素E组表现明显减轻。ELISA和Western-blot结果显示AR组鼻腔分泌物中IFN-γ、IL-17、IL-4和IL-5含量和表达水平明显高于对照组(P0.05),经γ-维生素E干预后,其含量明显降低(P0.05)。结论γ-维生素E减轻AR的临床表现可能与降低AR鼻腔分泌物中IFN-γ、IL-17、IL-4和IL-5的表达水平相关。     

NF-κB在变应性鼻炎发病中的调节作用

目的探讨变应性鼻炎(allergic rhinitis,AR)病理过程中TLR-NF-κB信号传导通路的调节机制及核因子-κB(NF-κB)对嗜酸性粒细胞聚集的调节作用。方法 50只大鼠随机分成A组(对照组)、B组(AR组)、C组(AR+PGN)、D组(AR+PGN+Pam3CSK4)、E组(AR+PGN+BEL),应用卵清蛋白建造AR模型,HE染色观察鼻黏膜形态改变并计数炎症细胞浸润数,ELISA法检测鼻腔清洗液IFN-γ、IL-4、嗜酸性粒细胞主要碱性蛋白(MBP)及IgE的含量,Real-Time PCR及Western blot法检测Toll样受体2(TLR2)、NF-κB的表达情况。结果 B组变应性损伤明显,鼻黏膜嗜酸性粒细胞浸润数多于其他组,IL-4、MBP及IgE的表达较A组增高(P0.05);C组鼻黏膜以中性粒细胞浸润为主,IFN-γ、TLR2及NF-κB的表达较B组增高(P0.05);D、E组炎症细胞浸润数、IL-4、MBP及IgE的表达较B组降低(P0.05)。结论调控TLR-NF-κB传导通路的表达即抑制NF-κB的过度表达,有利于调节变应性鼻炎病理过程中嗜酸性粒细胞的增值聚集。     

MiR-155-5p介导Treg/Th17免疫失衡致变应性鼻炎机制研究

为探讨miR-155-5p异常表达对变应性鼻炎(allergic rhinitis, AR)小鼠Th17/Treg免疫平衡的影响及机制,将6周龄BALB/c小鼠随机平均分成6组:A.对照组(Control)、B.模型组(AR)、C.miR-155-5p空载对照组(AR+Vector)、D.miR-155-5p过表达组(AR+miR-155-5p)、E.miR-155-5p阴性对照组(AR+NC)、F.miR-155-5p沉默组(AR+抗miR-155-5p抗体)。采用卵清蛋白(ovalbumin, OVA)诱导建立AR小鼠模型,鼻腔内给予miR-155-5p过表达或沉默慢病毒进行干预。实验末对各组小鼠过敏症状进行评分,HE染色观察鼻黏膜组织病理学变化,real-time PCR、Western blotting检测鼻黏膜组织中miR-155-5p、Foxp3、RORγt的表达水平,ELISA检测血清中IgE及炎性因子TGF-β1、IL-10、IL-17A、IL-6表达,流式细胞术检测外周血Treg和Th17百分比。结果发现,与对照组比较,模型组小鼠鼻炎症状评分明显增加,Foxp3、RORγt mRNA及蛋白表达水平明显升高,血清IgE、IL-17A、IL-6及Th17百分比显著上升,TGF-β1、IL-10及Treg百分比明显降低。MiR-155-5p过表达加重模型组小鼠鼻炎症状,同时促进IgE及Th17相关细胞因子的表达,抑制Treg相关细胞因子的表达,而miR-155-5p沉默后Treg/Th17免疫失衡得到缓解,小鼠鼻炎症状评分显著下降。MiR-155-5p可以显著加重AR小鼠的鼻炎症状,可能与影响Foxp3、RORγt表达,调节Treg/Th17免疫平衡有关。     

腹股沟皮下注射法免疫治疗小鼠变应性鼻炎的实验研究

目的:探讨腹股沟皮下注射法免疫治疗小鼠变应性鼻炎(Allergic rhinitis,AR)的可行性。方法:36只雌性BALB/c小鼠,6~8周龄,随机分为6组,每组6只,包括A空白组、A模型组、A治疗组、B空白组、B模型组和B治疗组(A为腹股沟皮下注射,B为颈背部皮下注射)。卵清蛋白(Ovalbumin,OVA)AR模型建立成功后治疗组用OVA进行免疫治疗,隔天一次,共15次。鼻腔连续激发第7天时,进行小鼠AR症状评分,鼻黏膜变色酸2R染色统计鼻黏膜嗜酸性粒细胞(Eosinophils,EOS)数,ELISA法检测血清OVA-s Ig E表达和鼻腔灌洗液中IL-4和IFN-γ表达,计算IFN-γ与IL-4比值。以SPSS 17.0软件进行数据分析。结果:治疗前,模型组和治疗组AR症状评分均大于5,治疗后,A治疗组AR症状评分小于5。A空白组、A模型组、A治疗组、B空白组、B模型组和B治疗组EOS数分别为0.78±0.31、21.60±2.90、10.43±2.56、0.83±0.46、22.44±3.39和23.40±4.24,A治疗组EOS数较A模型组明显减少,差异具有显著性(P0.05),B治疗组EOS数与B模型组比较,差异无显著性(P0.05);空白组OVA-s Ig E的表达为阴性,其余组为阳性;各组IFN-γ与IL-4比值分别为10.75±3.38、10.38±3.08、3.02±0.69、2.71±0.89、2.52±0.30和5.45±1.41,A治疗组IFN-γ与IL-4比值较A模型组明显升高,差异具有显著性意义(P0.05),B治疗组IFN-γ与IL-4比值与B模型组比较,差异无显著性(P0.05)。结论:腹股沟皮下注射免疫治疗小鼠变应性鼻炎效果好,周期短,操作简单,可行性好,是特异性免疫治疗小鼠变应性鼻炎的一种新方法。     

IL-17A抗体抑制变应性鼻炎鼠模型中IL-17A及RORγt并升高Foxp3 mRNA表达

目的：探讨IL-17A抗体对小鼠变应性鼻炎鼻黏膜中IL-17A、RORγt、Foxp3 mRNA水平的影响及意义。方法：以卵清蛋白致敏的Balb/c小鼠变应性鼻炎模型作为实验组,同期以生理盐水替代作为对照组,使用IL-17A抗体治疗作为治疗组。实时定量PCR方法检测三组小鼠鼻黏膜中IL-17A、RORγt、Foxp3mRNA的含量。结果：实验组中RORγt以及IL-17AmRNA水平均较对照组升高(P<0.05),治疗组中RORγt以及IL-17AmRNA的含量均低于实验组(P<0.05)。而实验组中Foxp3 mRNA水平较对照组下降(P<0.05),治疗组中Foxp3 mRNA的含量高于实验组(P<0.05)。结论：IL-17A抗体抑制变应性鼻炎鼠模型中IL-17A及RORγt mRNA水平并升高Foxp3 mRNA表达水平。     

地塞米松对IL-13诱导的大鼠鼻黏膜mCLCA3和Muc5ac表达的影响

目的:研究地塞米松对白细胞介素IL-13诱导的大鼠鼻黏膜mCLCA3和黏蛋白Muc5ac表达的影响,探讨其对变应性鼻炎黏液分泌的影响.方法:30只SD大鼠随机分成对照组、IL-13组、地塞米松组,用逆转录聚合酶链反应法(RT-PCR)和免疫组化法分别检测鼻黏膜mCLCA3 mRNA和Muc5ac蛋白的表达.结果:对照组大鼠鼻黏膜中mCLCA3 mRNA表达为0.000±0.000,IL-13组为0.319±0.121,两组比较差异有统计学意义(P＜0.05);地塞米松组为0.144±0.105,IL-13组为0.319 ±0.121,两组比较差异有统计学意义(P＜0.05);对照组大鼠鼻黏膜中Muc5ac蛋白的表达(0.300±0.145)与IL-13组(1.389±0.499)比较,差异有统计学意义(P＜0.05);地塞米松组Muc5ac蛋白的表达(0.901 ±0.390)较IL-13组(1.389±0.499)明显减少(P＜0.05).结论:IL-13能够上调mCLCA3 mRNA和Muc5ac在鼻黏膜的表达,地塞米松可下调mCLCA3 mRNA和黏蛋白Muc5ac表达.     

克鼻敏汤剂对实验性变应性鼻炎大鼠细胞因子水平的调节作用

研究中药复方克鼻敏汤剂对实验性变应性鼻炎大鼠的治疗作用,并初步探讨其机制。用卵清蛋白致敏大鼠制作变应性鼻炎动物模型,并经灌胃克鼻敏进行预防性治疗。观察变应性鼻炎大鼠行为学改变;组织病理学方法观察鼻黏膜改变;采用ELISA法检测血清总IgE、卵清蛋白特异性IgE和细胞因子IFN-γ、IL-2、IL-4及IL-5水平。结果发现与模型对照组比较,克鼻敏治疗组行为学积分、血清总IgE和特异性IgE含量,IL-4和IL-5水平均明显下降(P<0.01);血清IL-2和IFN-γ明显升高(P<0.01)。克鼻敏治疗组与药物对照组和正常对照组比较,IFN-γ、IL-4和IL-5水平均无显著性差异,但克鼻敏治疗组行为学积分、血清总IgE和特异性IgE含量及IL-2水平均高于药物对照组和正常对照组。克鼻敏治疗组与强的松治疗组比较各项观测指标均无显著性差异。研究结果提示中药复方克鼻敏汤剂可通过调节细胞因子表达,下调IgE的产生达到对实验性变应性鼻炎的治疗目的。     

Tim-3/galectin-9调控脂多糖诱导的人淋巴细胞中Th1/Th2细胞平衡

目的 探讨T细胞免疫球蛋白黏蛋白分子-3(Tim-3)/半乳糖凝集素-9(galectin-9)在脂多糖(LPS)诱导淋巴细胞炎性过程中对辅助性T细胞(Th)Th1/Th2细胞失衡中的作用。方法 收集脓毒症患者的外周血。分离外周血单个核细胞(PBMC),RT-qPCR检测Tim-3、galectin-9、T-bet和GATA3的表达；ELISA检测血清sTim-3、galectin-9、IFN-γ和IL-4的水平；分离的外周血单个核细胞体外培养，LPS刺激建立体外炎性模型；ELISA法检测细胞培养上清中Th1和Th2细胞相关细胞因子的水平；Western blot检测JAK2/STAT3相关因子的表达；Pearson相关系数分析Tim-3、galectin-9、IFN-γ和IL-4的相关性。结果 脓毒症患者外周血中Tim-3和galectin-9的mRNA表达增多，sTim-3、galectin-9、IL-4和GATA-3升高，IFN-γ和T-bet水平下降，磷酸化JAK2和STAT3升高(P<0.05)。阻断Tim-3表达后，IFN-γ水平升高，而IL-4的水平明显下降，磷酸化...     

Th9细胞在小鼠变应性鼻炎中的表达

目的探讨辅助性T细胞9(Th9)在变应性鼻炎小鼠鼻黏膜中的表达及意义。方法构建以卵清蛋白致敏的Balb/c小鼠变应性鼻炎模型作为实验组,同期使用生理盐水作为对照组。取2组小鼠鼻黏膜组织,使用流式细胞仪检测2组小鼠鼻黏膜中Th9细胞比例。结果 2组小鼠鼻黏膜中均有Th9细胞的存在,并且实验组中Th9的比例高于对照组[(1.265%±0.381%)vs(0.616%±0.041%)],差异具有统计学意义(P0.05)。结论首次明确变应性鼻炎鼻黏膜中Th9细胞的比例,为阐明AR的发病机制提供了新的理论依据。     

Rates of oxygen consumption and of anaerobic glycolysis in renal cortex and medulla of adult and new-born rats and guinea-pigs

1. Rates of oxygen uptake and of anaerobic glycolysis were estimated in slices from the renal cortex and medulla (a) of adult rats and guinea-pigs, (b) of new-born (1-, 5- and 21-day-old) rats and of guinea-pigs of 1, 12, 21, 24 and 120 hr age.2. In the adult rat, Q(O2) values for the cortex were 12.55 +/- 0.20 (22) and for the medulla: 8.56 +/- 0.17 (22) mul./hr.mg dry weight, while in the new-born rat (24 hr old) they were 10.99 +/- 0.46 (12) and 9.33 +/- 0.18 (9) mul./hr.mg dry weight respectively.3. Values for Q(CO2) (N2) (anaerobic glycolysis) in the 14 hr old new-born rat were in the renal cortex 9.65 +/- 0.35 (5) and in the medulla 7.39 +/- 0.43 (5) mul./hr.mg dry weight; while in the adult they were 2.25 +/- 0.08 (16) and 5.76 +/- 0.14 (16) mul./hr.mg dry weight, respectively.4. In the adult guinea-pig values for Q(CO2) (N2) were of the same order as in the adult rat, though the rate of O(2) uptake was for the cortex 8.12 +/- 0.22 (12) and for the medulla 5.02 +/- 0.23 (11) mul./hr.mg dry weight.5. Though the Q(O2) values in the renal cortex and medulla were smaller in the 1 hr old new-born guinea-pig, they were already increasing in the 12 hr old neonate.6. The results are discussed in the light of enzyme changes occurring during the process of maturation of the nephron as indicated by histochemical observations.     

Modes of reinforcement and gender and culture of experimenter and subject: effects of "alien" and external reinforcement for Japanese and American men and women

In this study, we examined external and "alien" reinforcement (ER and AR. respectively) as a factor in social learning, and studied the combined effects of culture and reinforcement mode. A female (Experiment 1) and a male (Experiment 2) experimenters conducted experimental sessions. Both men and women, who grew up in the same culture as the experimenter, participated and performed the experimental task. A three-way interaction effect of experimenter gender, culture, and reinforcement mode was found on task performance. And the effect was more pronounced for a Japanese experimenter. A female and a male experimenters conducted Experiments 3 and 4, respectively; however participants this time were men and women who grew up in different cultures than the experimenter. Results indicated that the pattern of the subject gender and reinforcement mode interaction effect, when the experimenter was Japanese with American subjects, was exactly opposite to that when the experimenter was American. These experiments showed that AR was as effective for social learning as ER, and that the cultural backgrounds of experimenter and subject influenced AR and ER effectiveness.     

Timing and intensity of early fevers and the development of allergies and asthma

BACKGROUND: Early childhood fevers appear to protect against later allergies and asthma. What is not known is the time in which fevers exert this effect and whether the degree of temperature increase is important. OBJECTIVE: We sought to examine the relationship between the time and degree of early fevers and later allergies and asthma. METHODS: Eight hundred thirty-five children from southeast Michigan were enrolled at birth. Clinic records from their first 2 years were abstracted for episodes of fever. At age 6 to 7 years, children underwent allergy testing. We examined fevers occurring within 6-month intervals in the first 2 years of life and outcomes at age 6 to 7 years. The primary outcome measures were allergic sensitization, asthma, asthma with allergic sensitization, and asthma without allergic sensitization. RESULTS: In the unadjusted analysis each episode of fever between 7 and 12 months of age was associated with a lower odds of allergic sensitization (odds ratio [OR], 0.71; 95% CI, 0.54-0.93) and asthma with allergic sensitization (OR, 0.43; 95% CI, 0.21-0.90) at age 6 to 7 years. Likewise, every 1 degrees C increase in the maximum temperature between 7 and 12 months was associated with a lower odds of allergic sensitization (OR, 0.77; 95% CI, 0.61-0.96) and asthma with allergic sensitization (OR, 0.62; 95% CI, 0.40-0.94). After adjusting for potential confounders, each episode of fever between 7 and 12 months was associated with a lower likelihood of asthma with allergic sensitization (adjusted OR, 0.33; 95% CI, 0.11-0.94) at age 6 to 7 years. CONCLUSIONS: Both the timing and intensity of childhood fevers appear to be important factors in the development of allergies and asthma.     

纳米水平分子成像与诊疗一体化研究进展与挑战

分子成像能以非侵入性的方式重现活体细胞的生理功能和生物学过程,提高疾病的早期和特异性诊断水平。纳米颗粒/材料具有物理性质可控性高、易于表面修饰、血液循环时间长和可功能化等优点,在疾病诊断与治疗中显示出巨大潜力。但如何阐明纳米材料多功能间的内在联系、解决其代谢及安全性等关键机制难题、实现纳米颗粒/材料多功能性到临床多功能...