益生菌对儿童中重度特应性皮炎的辅助治疗

目的探讨益生菌辅助治疗对儿童中重度特应性皮炎(AD)的临床疗效。方法选择于首都儿科研究所附属儿童医院皮肤科就诊的96例中重度AD患儿为研究对象。入选患儿随机分成两组,其中试验组(50例)患儿给予口服乳杆菌及我院皮肤科AD常规药(口服抗组胺药,外用糠酸莫米松乳膏、我院自制肤乐霜);对照组(46例)患儿仅给予我院皮肤科AD常规药。两组患儿均治疗3个月,治疗前后分别用SCORAD评分、CDLQI评分、DFIQ评分进行病情评估,同时检测两组患儿治疗前后的血清总IgE、IL2受体、IL4、嗜酸细胞阳离子蛋白(ECP)水平及Th1/Th2比值。结果治疗后,SCORAD评分显示实验组患儿总有效率为82.0%(41/50),对照组总有效率为52.2%(24/46),两者差异有统计学意义(χ²=6.448,P=0.011)。治疗后,试验组患儿SCORAD评分、CDLQI评分、DFIQ、总IgE水平低于对照组(t=4.656,4.189,3.097、2.278;P<0.001、<0.001、=0.003、=0.025),而IL2受体、IL4、ECP、Th1/Th2水平差异无统计学意义(均P>0.05)。结论中重度AD患儿在常规治疗上基础上补充益生菌,可显著降低血清IgE水平,减轻AD患儿的症状,提高疗效。     

益生菌对婴儿期特应性皮炎预防作用的系统评价

目的系统评价益生菌对婴儿期特应性皮炎的预防作用。方法计算机检索中英文数据库,检索时限从建库到2017年12月,收集益生菌对婴儿期特应性皮炎预防的临床随机对照试验的文献,由2名研究员筛选文献及质量评价,采用RevMan 5.3软件进行Meta分析。结果最终纳入质量较高的20个研究,共3 701例患儿。结果显示:益生菌组特应性皮炎发生率低于对照组,两组差异具有统计学意义(RR=0.74,95%CI:0.64~0.86,P0.01)。进一步亚组分析后发现仅产前母亲或产后婴儿口服益生菌对婴儿期特应性皮炎无预防作用(RR=0.91,95%CI:0.62~1.33,P=0.62;RR=0.91,95%CI:0.71~1.16,P=0.43),而在两者均服用益生菌的比较中发现单独服用乳酸菌类或与其他益生菌混合服用同安慰剂组比较均具有预防作用(RR=0.70,95%CI:0.52~0.94,P0.05;RR=0.65,95%CI:0.50~0.86,P0.01)。结论孕母及产后婴儿均补充益生菌对婴儿期特应性皮炎具有预防作用,单独服用临床常用的乳酸菌类或与其他益生菌混合服用同样具有保护作用,但由于国内外菌株、种群差异等诸多因素,其对婴儿期特应性皮炎的保护机制在国内尚需进一步研究。     

益生菌对特应性皮炎的防治及机制研究进展

特应性皮炎(atopic dermatitis, AD)是一种以反复发作和严重瘙痒为特征、发病率最高的过敏性皮肤病。AD的致病机制涉及遗传易感性、表皮屏障功能障碍、微生物组失调、免疫反应失衡以及环境等多个因素,而现有治疗用药副作用大、疗效欠佳。目前研究已发现肠道菌群尤其是益生菌在AD中起着重要作用。益生菌能够通过抑制病原菌、增强屏障功能、改善肠道环境和平衡Th1/Th2免疫应答等机制改善AD症状。本文综述了AD患者皮肤及肠道微生态特征,基于AD发病的致病机制和影响因素,系统阐明益生菌缓解AD的机制,以期为益生菌治疗AD及相关皮肤过敏性疾病提供理论支持。     

DNCB致敏/激发Nc/Nga小鼠特应性皮炎模型的建立

目的研究外用2,4-二硝基氯苯（DNCB）对Nc/Nga小鼠的致敏作用,探索建立特应性皮炎（AD）模型的方法。方法外用1%DNCB7周,间隔为1周,重复刺激并致敏7周大NC/Nga小鼠的双侧耳朵及背部皮肤。结果外用DNCB7周可以引起显著的炎症,并伴有高滴度的IgE和IL-4,利用HE染色进行组织病理分析,提示表皮炎性细胞明显增加。结论外用DNCB重复刺激Nc/Nga小鼠能产生AD样皮炎,可以作为研究AD病因及治疗AD的有效动物模型。     

布拉酵母菌联合左西替利嗪治疗小儿特应性皮炎的 临床疗效及安全性评价

摘要：目的 观察布拉氏酵母菌联合左西替利嗪口服液治疗小儿特应性皮炎的临床疗效及安全性。方法 随机将80例特应性皮炎患儿分为观察组和对照组各40例。对照组采用左西替利嗪口服液,2~5岁5 mL/次,1次/d：≥6岁 10 mL/次,1次/d。观察组在对照组基础上联合布拉氏酵母菌散剂,<3岁1袋 qd,≥3岁1袋,bid口服。2周1个疗程, 治疗结束 1个月后评价两组临床疗效及安全性。结果 总有效率：治疗组为90.00%,对照组为67.50%,两组比较差异有统计学意义（统计值,P＜0.05）。复发率：治疗组4例复发（10.00%）,对照组12例复发（30.00%）,两组比较差异有统计学意义（统计值,P＜0.05）。结论 布拉氏酵母菌联合左西替利嗪治疗小儿特应性皮炎临床疗效优于单用左西替利嗪,未见明显不良反应。     

基于微生物视角的“皮-肠”轴与特应性皮炎

特应性皮炎(atopic dermatitis,AD)是一种难治易复发皮肤病,由于病因复杂且患病率逐年增加,该病已经成为公共卫生领域关注的问题。随着高通量测序、元基因组学和代谢组学等技术的应用,发现AD的发生与发展与微生物群落息息相关,“微生物-皮-肠”轴及它们之间的串扰机制也逐渐被验证。“微生物-皮-肠”轴在过敏性皮肤炎症中扮演了重要角色。本文综述了“微生物-皮-肠”轴与AD的关系,及其可能交流的信号分子和潜在途径,重点关注了涉及益生菌、菌群移植和抗菌肽等微生物缓解AD的潜在机制,为靶向微生物群治疗过敏性皮肤炎症提供了一个新的视角。     

氟芬那酸丁酯软膏联合丁酸氢化可的松治疗儿童慢性湿疹及特应性皮炎的疗效

目的:探讨氟芬那酸丁酯软膏联合丁酸氢化可的松治疗儿童慢性湿疹及特应性皮炎的临床疗效。方法:选取我院诊治的110例慢性湿疹及特应性皮炎患儿作为研究对象,使用随机数字表法分为研究组和对照组(各55例),对照组单用丁酸氢可的松软膏治疗,研究组联合应用氟芬那酸丁酯软膏和丁酸氢化可的松软膏,比较两组患儿的治疗效果。结果:研究组治疗后的疾病积分显著低于对照组,P0.05;研究组的治疗总有效率为85.45%,对照组为63.64%,组间比较,差异具有统计学意义(P0.05)。结论:临床治疗儿童慢性湿疹和特应性皮炎,联合应用氟芬那酸丁酯软膏和丁酸氢化可的松,具有显著疗效,且安全性好,值得推广。     

凝结芽孢杆菌-乳果糖合生元对DSS诱导的溃疡性结肠炎小鼠肠道健康的影响

[目的]旨在探究凝结芽孢杆菌-乳果糖合生元对葡聚糖硫酸钠(dextran sodium sulfate,DSS)诱导的溃疡性结肠炎小鼠临床体征、肠道形态和肠道菌群结构的影响.[方法]选取24只初始体重为(22.96±1.87)g的7周龄雄性C57/BL6小鼠,随机分为4组,每组6只,即CON组、DSS组(连续5 d饮用...     

草鱼树突状细胞的分离鉴定及益生芽孢杆菌对其免疫功能的影响

为揭示草鱼(Ctenopharyngodon idellus)树突状细胞(Dendritic cells)的生物学特性及益生芽孢杆菌对其免疫功能的影响,研究通过草鱼体外细胞培养技术分离获得草鱼DCs,对其形态学特征、生物学功能及膜表面标记分子的表达进行了分析鉴定; RT-PCR检测了益生芽孢杆菌对草鱼DCs免疫相关细胞因子表达的影响。结果表明草鱼DCs具有典型的树突状形态,可有效地激活T淋巴细胞的增殖并具有迁移能力。LPS刺激可促进其成熟过程,显著提升膜表面标记分子CD80/86、CD83的表达。这表明草鱼DCs和哺乳动物DCs在形态和功能上具有高度相似性。RT-PCR结果显示,在体外条件下使用紫外照射灭活的益生枯草芽孢杆菌对草鱼DCs进行刺激后,抗炎性因子IL-4, IL-10的表达水平显著提升(P<0.05),并在12h时达到峰值。这表明枯草芽孢杆菌可通过促进树突状细胞分泌抗炎性因子来影响其免疫功能。以上结果为进一步研究鱼类树突状细胞的生物学特性及益生芽孢杆菌对其免疫功能的影响提供了重要的依据。     

不同严重程度儿童特应性皮炎血清半乳糖凝集素-10及炎症因子的表达水平及其与皮肤感染的相关性分析

摘要 目的：分析不同严重程度儿童特应性皮炎血清半乳糖凝集素-10（Gal-10）及炎症因子的表达水平及其与皮肤感染的相关性。方法：选择我院自2022年1月至2023年1月收治的110例特应性皮炎患儿纳入观察组,根据特应性皮炎评分（SCORAD）,分为轻度组、中度组和重度组;另选110例健康体检儿童纳入对照组。检测所有入选者血清Gal-10、炎症因子（IL-4、IL-13、IL-31）水平及不同部位皮肤经皮失水量,分析血清Gal-10、IL-4、IL-13、IL-31与SCORAD评分、不同部位皮肤经皮失水量的关系,使用受试者工作特征曲线（ROC）分析血清Gal-10、炎症因子（IL-4、IL-13、IL-31）对皮肤感染的预测效能。结果：观察组血清Gal-10、IL-4、IL-13、IL-31水平均高于对照组（P＜0.05）;随着儿童特应性皮炎病情加重,患儿血清Gal-10、IL-4、IL-13、IL-31水平随之升高,血清Gal-10、IL-4、IL-13、IL-31水平在轻度组、中度组和重度组中差异显著（P＜0.05）;经Pearson相关性分析,特应性皮炎患儿血清Gal-10、IL-4、IL-13、IL-31水平与SCORAD评分,前壁伸侧、前壁屈侧、脸颊及胫前的经皮失水量呈正相关（P＜0.05）;在110例特应性皮炎患儿中,发生皮肤感染43例;经ROC曲线分析,血清Gal-10、IL-4、IL-13联合IL-31预测特应性皮炎患儿发生皮肤感染的敏感度为89.72%、特异度为56.97%、ROC曲线下面积（AUC）为0.921。结论：血清Gal-10、炎症因子（IL-4、IL-13、IL-31）水平与儿童特应性皮炎严重程度有显著相关性,对于评估其皮肤屏障功能和预测皮肤感染均具有一定作用,值得临床予以重视应用。     

Improvement in symbiotic efficiency of chickpea (Cicer arietinum) by coinoculation of Bacillus strains with Mesorhizobium sp. Cicer

Rhizobacteria belonging to Bacillus sp. were isolated from the rhizosphere of chickpea (Cicer arietinum). Ten Bacillus strains were studied for their antifungal activity, effect on seedling emergence and plant growth promotion. Two Bacillus strains CBS127 and CBS155 inhibited the growth of all the four pathogenic fungi tested on nutrient agar medium plates in vitro. Seed inoculation with different Bacillus strains showed stimulatory effect on root and shoot growth at 10 d of observation in comparison to control whereas four Bacillus strains CBS24, CBS127, CBS129 and CBS155 caused retardation of shoot growth at 10 d. Maximum nodule-promoting effect was observed with Bacillus strains CBS106, CBS127 and CBS155. The symbiotic effectiveness of Mesorhizobium sp. Cicer strain Ca181 was further improved on coinoculation with six Bacillus strains i.e. CBS9, CBS17, CBS20, CBS106, CBS127 and CBS155 at 80 d of plant growth under sterile conditions and shoot dry weight ratios increased 1.62 to 1.74 times those of Mesorhizobium-inoculated treatments, suggesting the usefulness of introduced rhizobacteria in improving crop productivity.     

Bacillus sp. mutant for improved biodegradation of Congo red: Random mutagenesis approach

This study presents the improved biodegradation of Congo red, a toxic azo dye, using mutant Bacillus sp. obtained by random mutagenesis of wild Bacillus sp. using UV and ethidium bromide. The mutants obtained were screened based on their decolorization performance and best mutants were selected for further studies. Better decolorization was observed in the initial Congo red concentration range 100–1000 mg/l for wild species whereas mutant strain was found to offer better decolorization up to 3000 mg/l. Mutant strain offered 12–30% reduction in time required for the complete decolorization by wild strain. The optimum pH and temperature were found to be 7.0 and 37 °C, respectively. Two efficient strains such as Bacillus sp. ACT 1 and Bacillus sp. ACT 2 were isolated from the various mutants obtained. Bacillus sp. ACT 2 showed improved enzymatic production and Bacillus sp. ACT 1 showed improved growth compared to wild strain. The enzyme responsible for the degradation was found to be azoreductase by SDS–PAGE and about 53% increased production of enzyme was achieved with mutant species. The experimental data were modeled using growth and substrate inhibition models.     

1株来自海洋的芽胞杆菌dhs-330转座突变库的构建及表面活性相关基因的克隆

为研究产生物表面活性剂的海洋芽胞杆菌dhs-330合成活性产物的分子机制,应用质粒pIC333介导的mini-Tn10转座子随机突变技术,构建了芽胞杆菌dhs-330的突变体库。通过表面活性测定和反向PCR克隆,从300个突变株中筛选出产表面活性剂水平提高的突变株2株,分别在ycsG和yvkC基因发生插入突变;表面活性降低的突变株4株,分别在fenC、yrkF、kinE和sigD基因发生插入突变。这些基因可能与芽胞杆菌dhs-330中表面活性剂的合成代谢和调控有关。     

Purification and characterization of a thermophilic alkaline xylanase from thermoalkaliphilic Bacillus sp. strain TAR-1

Thermoalkaliphilic Bacillus sp. strain TAR-1 isolated from soil produced an extracellular xylanase. The enzyme (xylanase R) was purified to homogeneity by ammonium sulfate fractionation and anion-exchange chromatography. The molecular mass of xylanase R was 40 kDa and the isoelectric point was 4.1. The enzyme was most active over the range of pH 5.0 to 10.0 at 50°C. The optimum temperatures for activity were 75°C at pH 7.0 and 70°C at pH 9.0. Xylanase R was stable up to 65°C at pH 9.0 for 30 min in the presence of xylan. Mercury(ll) ion at 1 mM concentration abolished all the xylanase activity. The predominant products of xylan-hydrolysate were xylobiose, xylotriose, and higher oligosaccharides, indicating that xylanase R was an endo-acting enzyme. Xylanase R had a K_m of 0.82 mg/ml and a V_max of 280 μmol min⁻¹ mg⁻¹ for xylan at 50°C and pH 9.0.     

花生侵脉新赤壳菌果腐病生防芽孢杆菌的分离鉴定及防病效果

【目的】为了分离鉴定对花生侵脉新赤壳菌果腐病病原菌Neocosmospora vasinfecta具有抑制作用的根际芽孢杆菌。【方法】利用平板稀释法从花生的根际土壤分离芽孢杆菌,再采用平板对峙法筛选出对N.vasinfecta具有抑制作用的根际芽孢杆菌,通过形态观察、生理生化特性和分子生物学相结合的多相分类方法对生防根际芽孢杆菌进行分类鉴定,检测脂肽类抗生素合成基因类型,并进行花生侵脉新赤壳菌果腐病的田间防治试验。【结果】从花生根际土壤中分离到28株芽孢杆菌,其中对花生果腐病病原菌具有明显抑制作用有8株。多相分类法结果显示2株为枯草芽孢杆菌(Bacillus subtilis),6株为解淀粉芽孢杆菌(B.amyloliquefaciens)。脂肽类抗生素合成基因检测显示,8株生防芽孢杆菌含有至少1种脂肽类抗生素,其中所有生防菌均含有丰原素B合成基因,推测这些芽孢杆菌对N.vasinfecta的抑制机制可能与脂肽类抗生素的合成相关。田间防病实验结果显示,B.amyloliquefaciens GF-3和GF-22制备的生物有机肥均能有效降低NPRP的发病指数,其防治效率分别为32.35%和79.41%,增产率分别为19.12%和25.85%。【结论】分离鉴定了2株对花生侵脉新赤壳菌果腐病具有明显防治效果的根际芽孢杆菌,这不仅为花生侵脉新赤壳菌果腐病的生防制剂研制提供了菌株,还为研究防治机理奠定了基础。     

Application of a thermostable glutamate racemase from Bacillus sp. SK-1 for the production of -phenylalanine in a multi-enzyme system

A gene encoding glutamate racemase (GluRA) was found in a thermophilic Bacillus strain named SK-1. The gene was cloned and expressed in Escherichia coli WM335, a -glutamate auxotroph. It consists of 792 bp with a start codon, TTG. The amino acid sequence deduced from the gene indicates that the GluRA has two cysteines and their surrounding regions are well conserved. The GluRA produced in the recombinant E. coli was purified to homogeneity by heat-treatment and Resource Q and Phenyl sepharose column chromatographies. The enzyme, which was determined to be a monomeric protein with a molecular weight of 29,000, did not require a cofactor such as pyridoxal 5′-phosphate, nicotinamide, or flavin for its activity. The enzyme was stable after incubation at 55 °C and retained 60% of its original activity after incubation at 60 °C. It was found to be stable in the region of pH 6.0–11.5. The thermostable GluRA was used as a catalyst in a multi-enzyme system composed of four enzyme reactions for the production of -phenylalanine. By running the multi-enzyme system for 35 h, 58 g l⁻¹ of -phenylalanine was produced with 100% of optical purity from equimolar amount of phenylpyruvate.     

Branchiibius cervicis sp. nov., a novel species isolated from patients with atopic dermatitis

Novel actinobacterial strains, PAGU 1247^T, PAGU 1251 and PAGU 1252, were isolated from the skin of atopic dermatitis patients and were characterized using a polyphasic approach. Phylogenetic analyses based on 16S rRNA gene sequences showed that these isolates were located within the family Dermacoccaceae. The most closely related species of PAGU 1247^T in phylogenetic terms was Branchiibius hedensis Mer 29717^T, with 16S rRNA gene sequence similarity of 99.6%, although the DNA–DNA relatedness value was less than 43.9%. Some biochemical traits, such as lipase (C14) and α-galactosidase activity, could distinguish these isolates from B. hedensis. Strain PAGU 1247^T contained iso-C_16:0 and brC_18:0 as the major fatty acids. The quinone system consisted of menaquinone MK-8(H₆ and H₄). The G + C content of the genomic DNA was 67.6 mol%. On the basis of its phenotypic properties and genetic distinctiveness, strains PAGU 1247^T, PAGU 1251 and PAGU 1252 represents a novel species of the genus Branchiibius, for which the name Branchiibius cervicis sp. nov. is proposed. The type strain is PAGU 1247^T (=NBRC 106593^T = DSM 24166^T).     

一株中度嗜盐芽孢杆菌(Bacillus sp. BZ-SZ-XJ39)的微生物学特性

【目的】了解一株来自新疆盐碱湖的中度嗜盐菌(Bacillus sp.BZ-SZ-XJ39)的微生物学特性,为进一步阐明该菌独特的盐适应机制奠定基础。【方法】通过16S rRNA基因序列分析、G+C mol%含量测定、细胞形态和菌落观察、培养条件确定、营养与生理生化指标测定以及细胞化学组分分析等描述该菌的生物特性。【结果】基于16S rRNA基因序列的同源性和系统发育树分析,菌株BZ-SZ-XJ39与Bacillus saliphilus 6AGT(序列相似性为97.5%),Bacillus daliensis DLS13T(96.5%),Bacillus luteus JC167T(96.2%),Bacillus chagannorensis CG-15T(95.5%)和Bacillus agaradhaerens DSM8721T(95.3%)有密切的亲缘关系。基因组DNA G+C含量为44.4 mol%±1.2 mol%(Tm)。菌株BZ-SZ-XJ39为好氧、短杆状、革兰氏阳性菌。菌落呈黄色、圆形凸起、表面光滑且边缘整齐。菌株生长盐度范围为0.5%–28%(最适8%),pH范围为5.5–9.5(最适pH 8.0),温度范围为4–41oC(最适33oC)。菌株BZ-SZ-XJ39合成的主要脂肪酸包括anteiso-C15:0(50.2%)和anteiso-C17:0(16.3%)。合成极性脂为磷脂(PL)、磷脂酰乙醇胺(PE)、磷脂酰甘油(PG)、二磷脂酰甘油(DPG)和磺酸基异鼠李糖基二脂酰基甘油(SQDG)。呼吸醌类型为甲基萘醌(MK-7)。菌株BZ-SZ-XJ39已经在中国普通微生物菌种保藏管理中心(CGMCC1.12936)和日本微生物菌种保藏中心(JCM30194)冻干保藏,在Gen Bank中的序列注册号为KP456019。【结论】基于BZ-SZ-XJ39菌株的遗传型、表型、生理生化以及化学组成特征,初步确定该菌为Bacillus属中的一个新成员。研究将为探索生命在高盐环境中存在的本质提供新素材,也可为生物技术的潜在应用提供新视角。     

Degradation of 2,4-dichlorophenol by Bacillus sp. isolated from an aeration pond in the Baikalsk pulp and paper mill (Russia)

A pure culture of 2,4-dichlorophenol (2,4-DCP)-degrading bacteria was isolated from a natural enrichment that had been adapted to chlorophenols in the aeration pond of the Baikalsk pulp and paper mill (Russia). The bacteria were identified by 16S rDNA intergenic region analysis, using PCR with universal primers. Comparative analysis of the 16S rDNA sequence (1545 bp) in the GenBank database revealed that these bacteria are related to Bacillus cereus GN1. Degradation of 2,4-DCP was studied using this culture in liquid medium under aerobic conditions, at initial concentrations of 20–560 μM 2,4-DCP. The 2,4-DCP degradation rates by B. cereus GN1 could be determined at concentrations up to 400 μM. However, higher concentrations of 2,4-DCP (560 μM) were inhibitory to cell growth.