THE EFFECTS OF MA-XING-GAN-SHI-TANG ON RESPIRATORY RESISTANCE AND AIRWAY LEUKOCYTE INFILTRATION IN ASTHMATIC GUINEA PIGS

Ma-Xing-Gan-Shi-Tang (MXGST), a traditional Chinese medicine, has been used in treatment of the bronchial asthma for several centuries. However, the therapeutic mechanisms of this Chinese medicine are still far from clear. To understand the mechanism of anti-asthmatic property of MXGST, a guinea pig model of allergic asthma was used to investigate the effects of MXGST on Dermatophagoides pteronyssinus-induced early and late asthmatic responses and airway inflammation, and examine direct β₂-adrenoceptor agonist activity in guinea-pig isolated trachea. Administration of MXGST (10 g/kg) extracts significantly inhibited the antigen induced immediate asthmatic responses (IAR) in actively sensitized guinea pig. MXGST caused concentration-dependent relaxation in strips of guinea pig trachea contracted with carbachol, and ICI-118551, a selective β₂-adrenoceptor antagonist, significantly inhibit the relaxation caused by MXGST. Furthermore, examination of bronchoalveolar lavage fluid (BALF) revealed that MXGST significantly inhibited the increase in neutrophil in the airway at 1, 6 and 24 hr after antigen challenge. Histopathologic examination results showed that MXGST suppressed the neutrophil infiltration into lung tissue. In conclusion, we suggest that the anti-asthmatic effects of MXGST are mainly due to its stimulation of β₂-adrenoceptors on bronchial smooth muscle and its anti-inflammatory ability to inhibit the neutrophil into the airway. The precise mechanism of action of MXGST in asthma remains to be elucidated.     

被动皮肤过敏反应大鼠肠系膜淋巴管的变化

目的观察被动皮肤过敏反应（PCA）大鼠肠系膜淋巴微循环的变化，探讨Ⅰ型变态反应性皮肤病的淋巴机制。方法清洁级雄性Wistar大鼠随机分为对照组、PCA1组、PCA2组、PCA3组。用卵清蛋白（OVA）作抗原制备抗血清，分别于大鼠背部脊柱两侧去毛处皮内注射5％、10％、20％抗血清，48h后再用10％OVA攻击致敏，复制PCA模型；对照组用生理盐水代替。各组动物分别于OVA攻击前10min及攻击后10min、20min、30min时测量肠系膜淋巴管口径，计算肠系膜淋巴管收缩性指数IndexⅠ、IndexⅡ、LD Index。结果成功复制了大鼠PCA模型。在OVA攻击前肠系膜淋巴微循环的各项指标均无统计学差异。在OVA攻击后30min，肠系膜淋巴管口径扩张、收缩频率增强、收缩性指数增加。结论PCA发生后，淋巴回流动力明显增强，可能是导致变态反应性皮肤病皮损区淋巴细胞浸润的重要因素之一。     

组胺激发试验对应变性鼻炎鼻粘膜微血管的影响

目的 研究组胺对应变性鼻患者鼻粘膜微血管的影响。方法 变应性鼻炎组28例,健康人组21例,用人体鼻粘膜微循环显微镜观察两组胺激发试验前后鼻粘膜微血管的管径、形态和管襻数的变化。结果 应变性鼻炎组组胺激发式试验后鼻粘膜微血管管径较激发前增粗,管襻数增加,鼻粘膜微血管形态以灯丝状以混合状增多。健康人组组胺激发试验前后鼻粘膜微血管管径、管襻数以形态均无变化。结论 变应性鼻粘膜微血管对组胺的反应性进行较健康人高,组胺对变应性鼻炎患者鼻粘膜容量血管和阻力血管均有舒张作用。     

Effect of Phlomis umbrosa Root on Mast Cell-Dependent Immediate-Type Allergic Reactions by Anal Therapy

The effect of aqueous extract of Phlomis umbrosa Turcz. (Labiatae) root (PUAE) on mast cell-dependent immediate-type allergic reaction by anal therapy was investigated. PUAE (0.01 to 1 g/kg) dose-dependently inhibited systemic anaphylaxis induced by compound 48/80 in mice. When PUAE was pretreated at the same concentrations with systemic anaphylaxis, the plasma histamine levels were reduced in a dose-dependent manner. PUAE (0.1 and 1 g/kg) also significantly inhibited local anaphylaxis activated by anti-DNP IgE. PUAE (0.001 to 1 mg/mL) dose-dependently inhibited the histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. The level of cyclic AMP (cAMP) in human mast cells (HMC-1 cells) when PUAE (1 mg/mL) was added, transiently and significantly increased compared with that of basal cells. In addition, PUAE (0.1 and 1 mg/mL) inhibited the secretion of tumor necrosis factor-α (TNF-α), interleukin (IL)-6 in phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187-stimulated HMC-1 cells. These results provide evidence that anal therapy of PUAE may be beneficial in the treatment of allergic diseases.     

Effect of foot shock stress on the interferon-gamma production of murine intestinal intraepithelial lymphocytes

To investigate the effect of stress on interferon (IFN)-γ production by intestinal intraepithelial lymphocytes (IEL), we exposed male C3H/HeN mice to electric foot shock for 30 min a day for 5 consecutive days. Immediately after the final foot shock stress, IEL from small intestine were isolated by Percoll density gradient. The stress induced a marked suppression of IFN-γ production by IEL stimulated with immobilized anti-CD3 mAb and a marked decrease in the proportion of IFN-γ-producing CD3⁺ IEL or αβTCR⁺ IEL stimulated with PMA + ionomycin. The αβTCR⁺ subset was the major cause of stress-induced suppression of IFN-γ production by IEL. Glucocorticoid induced the suppression of IFN-γ production by IEL in vitro, which was reversed by mifepristone (RU486), a glucocorticoid receptor antagonist. In vivo administration of RU486 reversed the stress-induced suppression of IFN-γ production by IEL. In conclusion, repeated foot shock stress suppressed IFN-γ production of IEL by stress-induced elevation of endogenous glucocorticoid. Substantial suppression of the αβTCR⁺ subset was the major cause of the suppression.     

Effect of Amino Acid Polymorphisms of House Dust Mite Der p 2 Variants on Allergic Sensitization

Purpose

The sequence variations of the Der p 2 allergen of Dermatophagoides pteronyssinus diverge along 2 pathways with particular amino acid substitutions at positions 40,47,111, and 114. The environmental prevalence and IgE binding to Der p 2 variants differ among regions. To compare IgE binding to Der p 2 variants between sera from Bangkok, Thailand and Perth, Western Australia with different variants and to determine the variant-specificity of antibodies induced by vaccination with recombinant variants.

Methods

The structures of recombinant variants produced in yeast were compared by circular dichroism and 1-anilinonaphthalene 8-sulfonic acid staining of their lipid-binding cavity. Sera from subjects in Bangkok and Perth where different variants are found were compared by the affinity (IC₅₀) of IgE cross-reactivity to different variants and by direct IgE binding. Mice were immunized with the variants Der p 2.0101 and Der p 2.0110, and their IgG binding to Der p 2.0103, 2.0104, and 2.0109 was measured.

Results

The secondary structures of the recombinant variants resembled the natural allergen but with differences in ANS binding. The IC₅₀ of Der p 2.0101 required 7-fold higher concentrations to inhibit IgE binding to the high-IgE-binding Der p 2.0104 than for homologous inhibition in sera from Bangkok where it is absent, while in sera from Perth that have both variants the IC₅₀ was the same and low. Reciprocal results were obtained for Der p 2.0110 not found in Perth. Direct binding revealed that Der p 2.0104 was best for detecting IgE in both regions, followed by Der p 2.0101 with binding to other variants showing larger differences. Mouse anti-Der p 2.0101 antibodies had a high affinity of cross-reactivity but bound poorly to other variants.

Conclusions

The affinity of IgE antibody cross-reactivity, the direct IgE binding, and the specificities of antibodies induced by vaccination show that measures of allergic sensitization and therapeutic strategies could be optimized with knowledge of Der p 2 variants.     

Activation of cytotoxic T lymphocyte responses and attenuation of tumor growth in vivo by Andrographis paniculata extract and andrographolide

The stimulatory effect of Andrographis paniculata extract and andrographolide on cytotoxic T lymphocyte (CTL) production was determined in BALB/c mice by Winn's neutralization assay using CTL sensitive EL4 thymoma cells as target cell. Extract and andrographolide showed a significant increase in CTL production in both the in vivo and in vitro models. The survival time of EL4 cells alone in animals was only 27.1 days and it was increased to 51.1 and 44.5 days in extract- and andrographolide treated animals with percentage increase in life span (%ILS) of 88.5 and 64.2, respectively. The survival rate of animals administered EL4 cells incubated with alloimmunized spleen cells (effector cells) from normal BALB/c mice was 35.8 (%ILS 32.1). When this group was treated with 10 doses of extract and andrographolide the life span was further increased to 52.1 days (%ILS 92.2 ) and 48.1 days (%ILS 77.4). Survival days of animal carrying EL4 cells incubated with alloimmunized spleen cells (effector cells) from extract and andrographolide treated animals were 55.5 and 50.3 days respectively. When these animals continued with extract and andrographolide treatment for 10 days their life spans were significantly increased to 62 and 53.8 days, respectively. The level of cytokines such as Interlevkin (IL)-2 and Interferon (IFN)-γ also was enhanced in these animals when they were treated with extract and andrographolide. This study demonstrated that A. paniculata extract and andrographolide stimulate the CTL production through enhanced secretion of IL-2 and IFN-γ by T cells and thereby inhibit the tumor growth.     

CD26 expression on CD4+T cells in patients with cutaneous leishmaniasis

Surrogate marker(s) of protection in human leishmaniasis is not well defined. In this study, T helper 1 (Th1) and Th2 cytokine profiles and CD26 expression on CD4(+) T cells in peripheral blood mononuclear cells of patients with healing or non-healing forms of cutaneous leishmaniasis (CL) stimulated with Leishmania antigens were assessed. The level of interferon (IFN)-gamma production was significantly higher in patients with healing or non-healing forms of CL than in healthy controls, but it was not significantly different between the two patient groups. The level of interleukin-5 production was significantly higher in patients with the non-healing form of CL than in the two other groups. There was a significant increase in the level of CD26 expression on CD4(+) T cells in patients with healing (P < 0.001) or non-healing (P = 0.025) forms of CL compared with the control group, but no significant difference was seen between the two patient groups. A weak positive correlation was seen between IFN-gamma production and CD26 expression on CD4(+) T cells of patients with the healing form of lesion (r = 0.54, P = 0.008), but this correlation was not observed in patients with the non-healing form of CL (r = 0.53, P = 0.078). Surface CD26 is not correlated with the clinical manifestation of CL or IFN-gamma production. Therefore, CD26 is not a surrogate marker for IFN-gamma production in CL.     

小青龙汤加减治疗过敏性鼻炎的疗效分析

目的 对小青龙汤加减治疗过敏性鼻炎的临床疗效进行观察和分析。方法 柴教授2016年1月~2017年5月门诊治疗的100例过敏性鼻炎患者为研究对象,随机分为中药治疗组和西药对照组,每组50例。西药对照组以氯雷他定片治疗,中药治疗组以小青龙汤加减治疗,对两组患者进行对比分析。结果 小青龙汤加味治疗过敏性鼻炎有效率98.00%优于西药组78.00%,中药治疗组临床症状体征评分(2.458±1.037)分低于西药对照组(4.629±1.725)分,差异有统计学意义（P<0.05）。结论 小青龙汤加减治疗过敏性鼻炎过程中,可明显改善患者临床症状,提高患者生活质量,值得临床推广应用。     

Suppression of Immediate and Late Anti-IgE-Induced Skin Reactions by Topically Applied Alcohol/Onion Extract

In a double blind study, alcohol/onion extract (5% ethanol) was injected simultaneously with 20 IU and 200 IU rabbit anti-human-IgE intradermally in 12 adult volunteers (6 atopics, 6 non-atopics). Diameters of wheals and flares were measured 10 min after and compared with control sites challenged with 20 IU and 200 IU anti-IgE in a 5% ethanol solution. The skin sites were then treated epidermally with 45% alcohol/onion extract and 45% ethanol under occlusion. Diameters of late cutaneous reactions were measured hourly. Oedema formation was clinically estimated according to an arbitrary scale and skin thickness measured with a calliper. In the onion-treated skin sites the wheal areas were significantly reduced (20 IU: control: 108 +/- 53 mm2; onion 69 +/- 42 mm2, P less than 0.05; 200 IU anti-IgE: control: 152 +/- 25 mm2, onion: 138 +/- 26 mm2, P less than 0.02). The oedema formation during the late phase skin reaction was markedly depressed (P less than 0.005 at 2 h, P less than 0.01 at 4 and 6 h, P less than 0.02 at 8 h). The extent of late skin reactions was slightly, but not significantly reduced. Obviously, onions contain pharmacologically active substances with anti-inflammatory and/or allergic properties.     

护理干预对门诊雾化吸入过敏性鼻炎依从性及其疗效

目的探讨护理干预对门诊雾化吸入治疗过敏性鼻炎患者的依从性及疗效的影响。方法收集门诊治疗的过敏性鼻炎患者115例,按就诊单双号分为干预组和对照组。对照组单纯应用布地奈德混悬液雾化吸入治疗,干预组给予布地奈德混悬液雾化吸入同时由护士给予护理干预。两组均连续用药14天,比较干预前后两组患者治疗依从性及临床症状改善情况。结果干预组较对照组临床症状积分下降指数显著增加(t=-6.177,P0.05),治疗依从性总评分有显著差异(t=-2.83,P=0.01)。结论护理干预能提高门诊过敏性鼻炎患者治疗的依从性,促进过敏性鼻炎患者临床症状的改善,提高患者治疗的疗效。     

地氯雷他定联合盐酸氮卓斯汀鼻喷剂治疗过敏性鼻炎疗效分析

目的 观察地氯雷他定联合盐酸氮卓斯汀鼻喷剂治疗过敏性鼻炎患者的临床效果。方法 选取2017年4月~2018年4月来我院就诊的过敏性鼻炎患者114例。采用随机数字表法划分法分为观察组与对照组,每组57例。对照组给予盐酸卓斯汀鼻喷剂,观察组在对照组治疗的基础上口服地氯雷他定,两组均连续治疗1个月,比较两组疗效、临床症状缓解时间、不良反应情况及指标的检测结果,随访3、6个月的复发率。结果 治疗后观察组总有效率为96.50%,高于对照组的82.46%,统计学意义显著（P＜0.01）。观察组的临床症状评分喷嚏（0.46±0.13）分、流涕（1.03±0.16）分、鼻痒（0.65±0.21）分、鼻堵（0.65±0.23）分均低于对照组的（0.87±0.21）分、（2.45±0.39）分、（2.41±0.52）分、（2.03±0.42）分,差异有统计学意义（P＜0.05）;观察组治疗后血清IgE、IL-4、Th1/Th2、IFN-γ的改善程度均优于对照组,统计学意义显著（P＜0.01）;观察组3月末复发6例（10.53%）、6月末复发14例（24.56%）均低于对照组的19例（33.33%）、27例（47.37%）,统计学意义显著（P＜0.01）;两组不良反应比较,差异无统计学意义（P＞0.05）。结论 地氯雷他定与盐酸氮卓斯汀鼻喷剂治疗过敏性鼻炎,其联合使用的疗效更显著,作用时间长,不良反应少且复发率低。     

Effects of Products of Activated Immune Cells and Recombinant Cytokines on Spontaneous and Ionophore-Induced Acrosome Reaction

PROBLEM: The purpose of this study was to examine whether the products of activated immune cells influence spontaneous and ionophore-induced sperm acrosome reaction. METHOD: The spontaneous and ionophore-induced acrosome reaction were evaluated by staining with fluorescein isothiocyanate (FITC) Pisum sativam agglutinin after incubation in capacitating media supplemented with either supernatants from Con-A activated leukocyte cultures or human recombinant (r) IL-Iβ, TNF-α, and INF-γ. RESULTS: The supernatants from Con A-activated peripheral blood leukocyte cultures at 1:1 and 1:10 dilution significantly increased the rate of spontaneous acrosome reaction (P < 0.001 and P < 0.01). Along with displayed abnormally elevated levels of spontaneous acrosome loss, sperm cells showed an insufficient ability to undergo acrosome reaction in response to the ionophore treatment. Recombinant IL-Iβ at increasing concentrations from 30 to 3 × 10⁴ U/ml did not have an effect on spontaneous and ionophore-induced acrosome reaction. In contrast, spermatozoa that underwent capacitation in media with 7 × 10³, 7 × 10⁴, and 7 × 10⁵ U/ml of rINF-γ showed a significant increase in spontaneous and induced acrosome reaction compared to the control (P < 0.001). Recombinant TNF-α at concentrations of 3.5 × 10³ U/ml and 3.5 × 10⁴ U/ml significantly inhibited ionophore-induced acrosome reaction (P <0.001). Both rINF-γ and rTNF-α together revealed an effect on the acrosome reaction similar to Con-A generated supernatants (1:1 and 1:10 dilution) only at the highest concentrations. CONCLUSIONS: Some cases of infertility may result from a defective acrosome reaction (premature acrosome loss or insufficient acrosome response to the stimulants) caused by products of activated lymphocytes and macrophages that are released into the male and female reproductive tracts.     

中西医结合治疗变应性结膜炎的临床观察

目的 观察中西医结合治疗变应性结膜炎的临床疗效。方法 选择2015年8月~2017年8月我院收治的变应性结膜炎患者80例作为研究对象,随机分为观察组和对照组,各40例。两组患者均给予西药治疗,观察组在此基础上口服中药方剂治疗。比较两组疗效及随访6个月的复发率。结果 观察组总有效率为95.00%,高于对照组的67.50%,差异有统计学意义（P＜0.05）;术后随访6个月,观察组复发率为5.00%,对照组为22.50%,组间比较,差异有统计学意义（P＜0.05）。结论 西药结合中药治疗变应性结膜炎临床疗效显著,术后复发率低。     

The Influence of IgE on Cultured Human Mast Cells

Purpose

The mast cell plays a pivotal role in the human immune response. Crosslinking of 2 IgE molecules bound to the high affinity IgE receptor (FcεRI) on the surface of the mast cell results in mast cell degranulation and the release of several proinflammatory mediators. Patients with type-I allergy have increased levels of IgE in the blood compared to healthy individuals.

Methods

In a 6-week culture system of stem cells to human mast cells we investigated the effect of the concentration of IgE. The mast cells were cultured with different concentrations of IgE for the last 10 days of the maturation period. It was observed how the IgE concentration affects the histamine release, FcεRI density on the mast cell surface and the concentration of other mediators.

Results

A clear correlation between IgE concentration in culture medium and the release of histamine upon activation was observed. It showed a bell-shaped dose response curve, with maximal response around an IgE-concentration of 250 ng/mL. Furthermore, the sensitivity of the mast cells and surface density of FcεRI on mast cell surface was also influenced by the IgE concentration in the culture medium.

Conclusions

IgE in the culture medium during the last 10 days of mast cell maturation influences the release of the preformed mediator histamine after mast cell activation and the density of FcεRI on the mast cell surface. The release of the de novo synthetized mediator prostaglandin D₂ and the expression of chymase and tryptase are not influenced by IgE in culture medium.     

白介素17单克隆抗体对变应性鼻炎小鼠气道炎症的作用

目的 探讨白介素17单克隆抗体（IL-17mAb）的不同给予剂量及方式在变应性鼻炎小鼠气道炎症中的作用。方法 将48只小鼠采用随机数字表法分为A、 B、 C、 D、 E、 F组,每组8只。分别于第0、 7、 14 d将20 μg卵清蛋白（OVA）加2 mg铝佐剂腹腔注射处理A、C、D、E及F组小鼠,间隔7 d,第22天开始进行鼻腔激发,每天每侧鼻孔各给予OVA 10 μl（共500 μg）滴鼻,连续7 d。A、C、D、E组小鼠于每次OVA鼻腔激发前1 h分别给予生理盐水、100 ng IL-17mAb、500 ng IL-17mAb、5 μg IL-17mAb滴鼻,F组小鼠于每次OVA鼻腔激发前4 h给予5 μg IL-17mAb腹腔注射,B组小鼠于相同时间点给予等量生理盐水腹腔注射及滴鼻。所有小鼠于最后1次激发后评估鼻部症状学变化,Diff-Quik染色观察鼻腔灌洗液（NLF）中嗜酸性粒细胞浸润情况,ELISA方法检测血清及NLF中IL-6、IL-10水平,鼻黏膜组织行甲苯胺蓝染色观察肥大细胞。结果 ４周末A组所有小鼠症状学评分均＞5分,提示造模成功。F组小鼠的挠鼻及喷嚏次数均少于A组（P＜0.05）;F组小鼠NLF中嗜酸性粒细胞数、血清IL-6水平低于A组,血清及NLF中IL-10水平均高于A组（P＜0.05）;E组小鼠血清中IL-10水平高于A组（P＜0.05）;A组小鼠鼻黏膜组织中肥大细胞数多于B组,统计学意义显著（P＜0.01）;F组小鼠鼻黏膜组织中肥大细胞数少于A组,但差异无统计学意义（P＞0.05）;F组小鼠鼻黏膜组织中肥大细胞数与B组比较,差异无统计学意义（P＞0.05）。结论 高剂量的（5 μg）IL-17mAb腹腔注射处于激发阶段的变应性鼻炎小鼠促使小鼠变应性鼻炎症状明显减轻,鼻腔灌洗液嗜酸性粒细胞减少。促使变应性鼻炎小鼠血清中IL-6表达降低,血清中及鼻腔灌洗液中IL-10表达升高,因此推测这些细胞因子的变化可能抑制Th17/促进Treg的分化,进而对变态反应产生抑制作用。     

孟鲁司特联合布地奈德治疗过敏性鼻炎的疗效及其对患者tIgE与sIgE的影响

目的研究孟鲁司特联合布地奈德治疗过敏性鼻炎的临床疗效和对患者tIgE与sIgE的影响。方法选取我院2011年7月到2015年5月间呼吸科及耳鼻喉科收治的160例过敏性鼻炎患者;随机分为观察组80例和对照组80例,观察组给予孟鲁司特联合布地奈德进行治疗,对照组单纯给予布地奈德进行治疗。比较两组患者治疗前后临床症状评分,同事对比两组治疗前后血清总免疫球蛋白E(TIg E)和特异性免疫球蛋白E(SIg E)浓度。结果观察组和对照组治疗前症状评分分别为11.21±3.46分、11.22±3.29分,治疗后观察组和对照组临床症状评分分别为1.96±0.79分、3.96±0.93分。两组间治疗前症状评分差异无统计学意义(P>0.05),治疗后均明显下降,而观察组下降更加明显,差异其有统计学意义(t=3.23,P<0.05)。观察组和对照组治疗前TIg E、SIg E水平分别为469.25±36.92k U/L、486.32±32.26k U/L、29.96±16.92k U/L、29.42±17.26 k U/L,差异无统计学意义(P>0.05);治疗后分别为249.35±27.13KU/L、362.28±35.18k U/L、12.35±7.13k U/L、14.28±6.18k U/L,两组间治疗后差异有统计学意义(P<0.05)。结论孟鲁司特联合布地奈德能够明显减轻过敏性鼻炎患者气道炎症反应,同时降低由Ig E介导的I型变态气道反应炎症,降低血清总免疫球蛋白E(TIg E)和特异性免疫球蛋白E(SIg E)浓度,是一种高效的治疗方法,值得在临床中推广使用。