支气管哮喘患者疾病认知状况调查及控制水平的影响因素分析

目的:调查支气管哮喘患者疾病认知状况,并分析控制水平的影响因素。方法:选取2018年7月~2020年7月期间贵州医科大学附属医院诊治的支气管哮喘患者100例,采用面对面问卷调查的方式调查所有患者疾病认知状况。采用哮喘控制测试(ACT)对患者哮喘控制水平进行评估。根据ACT结果将患者分为哮喘未控制组(n=57)和哮喘控制组(n=43)。分析哮喘控制水平的影响因素。结果:支气管哮喘患者对疾病认知相关问题的回答正确率均在60%以上,但仅有12%的患者使用过峰流速仪。本研究中100例患者均完成ACT,其中完全控制17例,控制良好26例,未控制57例,分别占比17.00%、26.00%、57.00%,哮喘控制率为43.00%。由单因素分析显示,支气管哮喘患者的哮喘控制水平与性别、家庭月收入、文化程度、家族史、吸烟史、居住处是否空气污染、病程、哮喘用药依从性、使用吸入性糖皮质激素治疗、抑郁情况、焦虑情况有关(P<0.05)。多因素Logistic回归分析显示:焦虑情况、抑郁情况、居住处空气污染、吸烟史是支气管哮喘患者哮喘控制水平的危险因素,而哮喘用药依从性、使用吸入性糖皮质激素治疗是支气管哮喘患者哮喘控制水平的保护因素(P<0.05)。结论:支气管哮喘患者对疾病有一定的正确认知,但仍未达到理想状态。哮喘控制水平受多种因素影响,可根据相关影响因素做出针对性的干预措施,以改善支气管哮喘控制水平。     

Lovastatin inhibits bronchial hyperresponsiveness by reducing RhoA signaling in rat allergic asthma

Recent studies revealed an importance of a monomeric GTP-binding protein, RhoA, in contraction of bronchial smooth muscle (BSM). RhoA and its downstream have been proposed as a new target for the treatment of airway hyperresponsiveness in asthma. Statins are known to inhibit the functional activation of RhoA via the depletion of geranylgeranylpyrophosphate. To determine the beneficial effects of statins on the airway hyperresponsiveness in allergic bronchial asthma, we investigated the effects of systemic treatment with lovastatin on the augmented BSM contraction and activation of RhoA in rats with allergic bronchial asthma. Rats were sensitized and repeatedly challenged with 2,4-dinitrophenylated Ascaris suum antigen. Animals were also treated with lovastatin (4 mg kg(-1) day(-1) ip) once a day before and during the antigen inhalation period. Repeated antigen inhalation caused a marked BSM hyperresponsiveness to ACh with the increased expression and translocation of RhoA. Lovastatin treatments significantly attenuated both the augmented contraction and RhoA translocation to the plasma membrane. Lovastatin also reduced the increased cell number in bronchoalveolar lavage fluids and histological changes induced by antigen exposure, whereas the levels of immunoglobulin E in sera and interleukins-4, -6, and -13 in bronchoalveolar lavage fluids were not significantly changed. These findings suggest that lovastatin ameliorates antigen-induced BSM hyperresponsiveness, an important factor of airway hyperresponsiveness in allergic asthmatics, probably by reducing the RhoA-mediated signaling.     

Phylogenetic analysis and heterologous expression of surface layer protein SlpC of Bacillus sphaericus C3-41

The surface layer protein encoding genes from five mosquito-pathogenic Bacillus sphaericus isolates were amplified and sequenced. Negative staining of the S-layer protein extracted from the cell wall of wild-type B. sphaericus C3-41 was prepared. It showed a flat-sheet crystal lattice structure. Two genes encoding the entire and N-terminally truncated S-layer protein (slpC and DeltaslpC respectively), were ligated into plasmid pET28a and expressed in Escherichia coli. SDS-PAGE revealed that about 130 KD and 110 KD proteins could be expressed in the cytoplasm of recombinant E. coli BL21(pET28a/slpC) and E. coli BL21(pET28a/DeltaslpC) respectively. Furthermore, an intracellular sheet-like or fingerprint-shape structure was investigated in two recombinant strains, which expressed SlpC and DeltaSlpC protein respectively, by ultrathin microscopy study, but bioassay results suggested that the S-layer protein of wild B. sphaericus C3-41 and recombinant E. coli BL21 (pET28a/slpC) have no direct toxicity against mosquito larvae. These results should provide information for further understanding of the function of S-layer protein of pathogenic B. sphaericus.     

球形芽胞杆菌C3-41磷酸果糖激酶的克隆、表达及基本生物活性

[目的]球形芽孢杆菌缺乏EMP、HMP、ED途径的关键酶,如磷酸果糖激酶等被认为是其不能以糖类物质进行生长的主要原因.杀蚊球形芽孢杆菌C3-41全基因组序列分析表明,在染色体DNA上存在的磷酸果糖激酶基因pfk,为了进一步分析球形芽孢杆菌糖酵解途径,进一步确定磷酸果糖激酶在糖酵解途径中的功能.[方法]通过pfk基因在球形芽孢杆菌菌株中的Southern-blot拷贝数鉴定,在C3-41pfk基因克隆的基础上进行pfk基因在大肠杆菌中的融合表达、序列分析和序列比对等方法进行研究.[结果]证明了球形芽孢杆菌pfk基因由960 bp核苷酸组成,表达42 kDa的PFK融合蛋白,有保守的底物结合域和ATP结合域,同时pfk基因重组表达质粒可以回复大肠杆菌pfk缺陷型菌株DFl020代谢糖的能力.[结论]杀蚊球形芽孢杆菌C3-41的pfk表达产物具有磷酸果糖激酶活性,为今后深入研究球形芽孢杆菌产能代谢机理奠定了基础.     

miR-1165-3p、miR-145水平在支气管哮喘患者中的表达及其临床意义

摘要 目的：探讨微小RNA（MicroRNA,miR）-1165-3p、miR-145水平在支气管哮喘患者中的表达及其临床意义。方法：收集2021年1月-2022年3月中国人民解放军总医院第六医学中心62例支气管哮喘患者作为研究组,其中轻度急性发作27例,中度急性发作22例,重度急性发作13例。另收集同时期、同年龄段30例健康体检者作为对照组。采用实时荧光定量PCR（RT-PCR）检测各组血清miR-1165-3p、miR-145表达水平。采用Spearman相关分析不同程度支气管哮喘患者与血清miR-1165-3p、miR-145之间的相关性。通过受试者工作特征（ROC）分析血清miR-1165-3p、miR-145表达水平对不同程度支气管哮喘的诊断效能。结果：与对照组相比,研究组中白细胞介素-6（IL-6）、嗜酸性粒细胞、总免疫球蛋白E（IgE）水平显著升高,第1秒用力呼气容积（FEV₁）占预测值百分比（FEV₁%）则显著降低,差异均有统计学意义（P＜0.05）。不同严重程度支气管哮喘患者（轻度、中度、重度）血清miR-1165-3p、miR-145表达水平均高于健康对照组,支气管哮喘越严重,其表达水平越高,且组间、组内比较差异均有统计学意义（P＜0.05）。Spearman相关分析显示,miR-1165-3p、miR-145、IL-6表达水平与哮喘严重程度呈正相关（P＜0.05）,FEV₁%与哮喘严重程度呈负相关（P＜0.05）,嗜酸性粒细胞、总IgE与哮喘严重程度无相关性（P＞0.05）。对轻度、中度、重度急性支气管哮喘发作的诊断效能显示：血清miR-1165-3p的曲线下面积（AUC）（0.95CI）分别为3.085（0.326～29.221）、0.712（0.611～0.829）、0.755（0.602～0.948）。血清miR-145的AUC（0.95CI）分别为0.833（0.708～0.979）、0.754（0.590～0.964）、0.816（0.671～0.993）。结论：血清miR-1165-3p、miR-145表达水平具有较高的诊断效能,支气管哮喘越严重,诊断的特异性越高,可作为支气管哮喘严重程度的无创诊断指标。     

支气管哮喘遗传因子研究

研究谷胱甘肽-S-转移酶 （glutathione S-transferase, GST）M1和T1基因多态性与支气管哮喘（asthma bronchial）的关系。采取聚合酶链反应对60名支气管哮喘患者和60名正常对照进行了GSTM1和GSTT1基因非缺失（+）和缺失（0）等位基因分布频率研究。结果表明,与对照组相比,支气管哮喘患者GSTM1基因缺失的纯合子（0/0）频率（81.2%）显著升高（χ2=32.46,P<0.001;wχ2=28.75,P<0.001）。对于GSTT1也得到类似资料。而支气管哮喘患者GSTT1基因缺失等位基因（0/0）频率（71.7%）比对照组（11.7%）显著升高（χ2=26.72,P<0.001;wχ2=35.75,P<0.001）。表明GSTM1、GSTT1缺失等位基因纯合性在哮喘患者中最有特征性的。GSTM1 0/0、GSTT1 0/0结合的频率患者组为61.7%,对照组仅为1.7%（χ2=27.3,P<0.001）。提示GSTM1和GSTT1基因多态性与哮喘有显著性关联,两个基因的突变可以被视为发生支气管哮喘遗传风险因子。     

STAT6 ASODN对哮喘小鼠脾淋巴细胞影响的实验研究

目的研究STAT6反义寡核苷酸对哮喘小鼠脾淋巴细胞的影响作用。方法实验细胞分组:正常鼠空白组(A组)、正常鼠OVA组(B组)、哮喘空白组(C组)、哮喘OVA组(D组)、哮喘治疗组(E组)。正常设计并人工合成一段互补于小鼠STAT6 mRNA翻译起始区271-290的反义寡核苷酸片段,全链硫代修饰。用卵白蛋白和氢氧化铝复制哮喘模型,用淋巴细胞分离液分离脾淋巴细胞,进行体外培养并导入由阳离子脂质体转染剂Geneshuttle携带的反义寡核苷酸,观察反义寡核苷酸的转染对脾淋巴细胞STAT6蛋白表达水平及细胞培养上清中IL-4分泌水平的影响。免疫细胞化学观察脾淋巴细胞中STAT6蛋白的表达水平,同时采用酶联免疫吸附(ELISA)法测定脾细胞培养上清液中IL-4的浓度。结果D组细胞STAT6蛋白表达明显高于其余各组,均具有显著性差异(P均<0.01),STAT6 ASODN转染后,E组细胞该蛋白的表达量明显下降(P<0.01);D组脾淋巴细胞培养上清中IL-4分泌水平明显高于其余各组,均具有显著性差异(P均<0.01);STAT6 ASODN转染后,E组培养上清中IL-4分泌水平显著低于D组(P<0.01)。结论STAT6 ASODN可特异性抑制哮喘鼠脾淋巴细胞中STAT6蛋白的表达,并可特异性抑制脾淋巴细胞中IL-4的分泌,为反义基因技术治疗哮喘提供了依据。     

New high-toxicity mosquitocidal strains of Bacillus sphaericus lacking a 100-kilodalton-toxin gene.

Five new high-toxicity mosquitocidal strains of Bacillus sphaericus were isolated in Singapore. They all belong to phage group 8 and have binary toxin (51.4- plus 41.9-kDa) genes located on the chromosome but lack a 100-kDa-toxin gene. These strains of B. sphaericus constitute a new subgroup, as only two weakly toxic strains in phage group 8 have previously been described and all the known high-toxicity strains have both binary toxin and 100-kDa-toxin genes.     

The influence of the bronchodilatation test on the duration of forced expiratory tracheal noises in young men

The diagnostic efficiency of estimating the duration of forced expiratory noises under the conditions of bronchial obstruction has been shown. The objective of this study was to analyze the response of the forced expiratory noise duration to the bronchodilatation test with the β₂-agonist in the age- and genderhomogenous group of healthy volunteers and bronchial asthma patients selected as a model of variable bronchial obstruction. Two hundred and sixty young men (16–25 years old) were examined. It was shown that the prevailing type of response in bronchial asthma patients with spirometry confirmed bronchial obstruction was shortened forced expiratory noises. Furthermore, the degree of the shortening considerably depended on the severity of the background bronchial obstruction. The absence of a statistically significant response of the forced expiratory noise duration dominated among healthy volunteers (nonsmokers as well as smokers) and bronchial asthma patients without a spirometry confirmed bronchial obstruction. However, the shortened response occurred much more frequently in bronchial asthma patients than in healthy volunteers. The high specificity (86%) of the response as shortened forced expiratory noises to the β₂-agonist may be useful for diagnostics.     

孟鲁司特纳片联合沙美特罗替卡松粉吸入剂对儿童支气管哮喘的临床疗效

目的:探究孟鲁司特纳片联合沙美特罗替卡松粉吸入剂治疗儿童支气管哮喘的临床疗效并分析复发情况。方法:选取2011年9月至2012年12月在本院就诊并住院治疗的100例支气管哮喘患儿,随机分成的对照组以及研究组,各50例。对照组吸入沙美特罗替卡松粉吸入剂,研究组给予孟鲁司特纳片联用沙美特罗替卡松粉吸入剂,观察哮喘的复发和控制情况。结果:用药治疗后两组肺功能指标均有改善(P<0.05);研究组用药第3、6个月和停药6个月时患儿肺功能较对照组改善更明显,差异均有统计学意义(P<0.05);两组患儿治疗后临床控制率均有显著的改善,但仅研究组差异有统计学意义(P<0.05);研究组在用药3个月、6个月以及停药6个月三个阶段临床控制率均高于对照组,而在用药6个月以及停药6个月两个阶段复发率均低于对照组,差异均有统计学意义(P<0.05)。结论:孟鲁司特纳片联合沙美特罗替卡松粉吸入剂治疗儿童支气管哮喘疗效较单用沙美特罗替卡松粉吸入剂治疗效果更好,复发率低。     

Acute respiratory viral infections and bronchial asthma. Cellular and molecular aspects of the problem

In this review information on relationships between acute respiratory viral infections (ARVI) and attacks of bronchial asthma is presented. The mechanisms of the appearance of virus induced attacks and bronchial hyperreactiveness have been analyzed. The role of the immune response of the body as well as anti-inflammatory and chemotaxic cytokines in the development of bronchial obstruction and hyperreactiveness has been substantiated. The conclusion on the role of ARVI especially induced by respiratory syncytial virus and rhinoviruses, in the development of immune response facilitating allergic sensitization and provoking attacks of bronchial asthma has been made.     

Bronchial epithelial cells release IL-6, CXCL1 and CXCL8 upon mast cell interaction

Although mast cells have been found in increased numbers in bronchial epithelium in asthma patients, the pathogenic role of the interaction of mast cells with bronchial epithelial cells in the development of local inflammation in asthma is not well understood. In this study, primary human bronchial epithelial cells and a human mast cell line (HMC-1) were cultured either together or separately in the presence or absence of various signaling molecule inhibitors or dexamethasone. Cytokine IL-6, and chemokines including CXCL1 and CXCL8 in cell culture supernatant were assayed by enzyme-linked immunosorbent assay (ELISA), and the activity of mitogen-activated protein kinases (MAPKs), or nuclear factor-κB (NF-κB) in co-culture system was analyzed by ELISA. Co-culture of bronchial epithelial cells and mast cells induced a significant elevation of IL-6, CXCL1 and CXCL8 in bronchial epithelial cells, and both IL-17A and IL-17F could further enhance the release of these inflammatory mediators from co-culture. The induction of IL-6, CXCL1 and CXCL8 upon the interaction of bronchial epithelial cells with mast cells was mediated by MAPKs and NF-κB signaling pathways. These data indicate that the interaction of mast cells with bronchial epithelial cells may represent a crucial mechanism of regulating local inflammatory response in allergic asthma.     

Effect of inhalation therapy with beclomethasone dipropionate on unspecific bronchial hyperreactivity in patients with asthma]

An effect of beclomethasone dipropionate (Beclocort-Polfa) was investigated in the group of 24 patients with atopic and non-atopic bronchial asthma who have shown unspecific bronchial hyperreactivity to histamine during the periods of remission. The drug was administered in the form of aerosol in a daily dose of 1.0 mg for 4 weeks. Provocative histamine dose was established with Bronchoscreen device (Jaeger) thrice:prior to the treatment, after 1 week of placebo therapy, and after 1 month of beclomethasone administration. Statistically significant increase in histamine dose for provocation of bronchospasm (p < 0.01) being nearly twice higher than the baseline value has been noted in patients treated with beclomethasone dipropionate aerosol. It advocates such a treatment in unspecific bronchial hyperreactivity independently of the type of the bronchial asthma.     

Reduced Superoxide Dismutase in Lung Cells of Patients with Asthma

Lung cells recovered from symptomatic patients with asthma generate increased amounts of reactive oxygen species (ROS). Animal and in vitro studies indicate that ROS can reproduce many of the features of asthma. The ability of ROS to produce the clinical features of asthma may depend on an individual's lung antioxidant defenses. Patients with asthma are reported to have reduced antioxidant defenses in peripheral blood, but little is known about the antioxidant defenses of their lung cells. To define lung cell antioxidant defenses in asthma, the glutathione concentration and the glutathione reductase, glutathione peroxidase, catalase, and superoxide dismutase (SOD) activities were measured in cells recovered by bronchoalveolar lavage (BAL cells) and by bronchial brushing (bronchial epithelial cells, HBEC) from normal subjects and patients with asthma. Superoxide dismutase activity was reduced 25% in BAL cells (p < .05) and nearly 50% in HBEC (p < .02) from patients with asthma. Alterations in the other antioxidants were not identified. A direct relationship was found between airway reactivity to methacholine, measured as PC₂₀FEV₁, and HBEC SOD activity (r² = 89; p < .005), but not between airway reactivity and the other antioxidants. The finding of reduced SOD activity in lung cells of patients with asthma suggests that diminished SOD activity serves as a marker of the inflammation characterizing asthma. Alternatively, it may play a role in the development or severity of the disease. © 1997 Elsevier Science Inc.     

The effect of sodium nucleinate on allergic and immunological reactions

Experiments on immunized rabbits and guinea pigs indicated that sodium nucleinate (SN) was capable of weakening or entirely eliminating anaphylactic and skin reactions of delayed type hypersensitivity to repeated administration of staphyloanatoxin, APDT vaccine. The findings on patients with the infectious form of bronchial asthma and chronic rheumatism showed that sodium nucleinate attenuated reactions to the subcutaneous administration of staphylococcal and streptococcal allergens. The treatment of patients suffering from infectious-allergic bronchial asthma and rheumatism with SN resulted in the recovery of deficient T cells, T-suppressors, normalization of immunoglobulin concentrations. In children with acute glomerulonephritis sodium nucleinate normalized decreased T-suppressor cells and increased IgG and circulating immune complexes (CIC), resulting in a pronounced remission of disease. The mechanism of desensitization and elimination of CIC by SN has not been explored, however, the parameters of SN-induced immunomodulation are known rather completely. It is suggested that SN brings about accumulation in the cell of cyclic AMP which diminished membrane permeability, activates monoaminooxidase resulting in the degradation of histamine and other biogenic amines, enhances the synthesis of endogenous corticosteroids with their desensitizing properties. All these effects contribute to the elimination of delayed type hypersensitivity. The role of SN in the inhibition of delayed type hypersensitivity remains obscure.     

Determination of cysteinyl leukotrienes in exhaled breath condensate: method combining immunoseparation with LC-ESI-MS/MS

A rapid and precise method for the identification and quantification of cysteinyl leukotrienes (leukotriene C(4), leukotriene D(4) and leukotriene E(4)), essential markers of bronchial asthma, in exhaled breath condensate was developed. The protocol consists of immunoaffinity separation and a detection step, liquid chromatography combined with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). In particular, the selected reaction monitoring mode was used for its extremely high degree of selectivity and the stable-isotope-dilution assay for its high precision of quantification. The developed method was characterized with a high precision (≤ 7.7%, determined as RSD), an acceptable accuracy (90.4-93.7%, determined as recovery), a low limit of detection (≤ 2 pg/ml EBC) and a low limit of quantification (≤ 10 pg/ml EBC). It was compared to other simple, clinically appropriate combinations of pre-treatment methods (solid phase extraction and lyophilization) with LC/MS. Finally, the method (a combination of immunoaffinity separation with LC-MS) was successfully tested in a clinical study where a significant difference was found in the concentration levels of cysteinyl leukotrienes between patients with occupational bronchial asthma and healthy subjects.     

Pneumonia due to Enterobacter cancerogenus infection

Enterobacter cancerogenus (formerly known as CDC Enteric Group 19; synonym with Enterobacter taylorae) has rarely been associated with human infections, and little is known regarding the epidemiology and clinical significance of this organism. We describe a community-acquired pneumonia case in a 44-year-old female due to E. cancerogenus. Identification and antimicrobial susceptibility of the microorganism was performed by the automatized VITEK 2 Compact system (bioMerieux, France). The clinical case suggests that E. cancerogenus is a potentially pathogenic microorganism in determined circumstances; underlying diseases such as bronchial asthma, empiric antibiotic treatment, wounds, diagnostic, or therapeutic instruments.     

Action of Bacillus sphaericus endopeptidases on bacterial peptidoglycans and peptidoglycan fragments

Two endopeptidases have been characterized in Bacillus sphaericus 9602: a gamma-D-glutamyl-(L) meso-diaminopimelate endopeptidase (endopeptidase I) and a gamma-D-glutamyl-L-diaminoacid endopeptidase (endopeptidase II). They are active on the peptide moieties of some bacterial peptidoglycans. Their specificities have been studied on peptides or monomeric glycopeptides derived from peptidoglycans. Their study was attempted on dimeric and polymeric fragments of a E. coli radioactive peptidoglycan. Those compounds are specifically labelled on the meso-diaminopimelate residues and are listed below.     

喘息的"度"与支气管哮喘阶梯治疗

目的:探讨掌握喘息"度"与支气管哮喘阶梯治疗的关系。方法:充分掌握支气管哮喘分级治疗方案和支气管哮喘病情,将喘息"度"细致化,与分级治疗用药"度"密切结合起来,对于完全理想的控制哮喘将会起到极大的指导作用,对于升阶梯和降阶梯治疗都会起到重要而细致的指导作用。必须将平喘治疗措施置于患者全身病情变化及总体治疗之下,会取得更加理想的哮喘控制效果。而强化支气管哮喘患者教育在支气管哮喘理想治疗中占据极为重要的地位,甚至直接关系到哮喘控制的持久性和稳定性。应得到高度重视。结果:喘息"度"与支气管哮喘的发作程度密切相关,准确把握并分级十分重要。结论:准确把握喘息"度"并与支气管哮喘分级治疗方法结合,将对稳定平息支气管哮喘起到意想不到的效果。