Mycoplasma pneumoniae induces interleukin-8 production via the epidermal growth factor receptor pathway

In mycoplasmal pneumonia, the bronchi are histopathologically filled with polymorphonuclear leukocytes. The EGFR pathway is involved in IL-8 production. We investigated the contribution of the EGFR pathway to IL-8 production by bronchial epithelial cells (A549) stimulated with Mp-Ag. The IL-8 production by A549 cells stimulated with Mp-Ag was decreased by the addition of an EGFR kinase inhibitor or transfection with small interfering RNA against EGFR. The levels of epiregulin mRNA in A549 cells were increased by stimulation with Mp-Ag. In conclusion, the EFGR pathway participates in IL-8 production by bronchial epithelial cells stimulated with Mp-Ag.     

Efficiency of Plasmocin™ on various mammalian cell lines infected by mollicutes in comparison with commonly used antibiotics in cell culture: a local experience

Mycoplasma contamination is a deleterious event for cell culture laboratories. Plasmocin™ is used to prevent and eradicate mycoplasma infections from cell. In this study, 80 different mammalian cell lines from various sources; human, monkey, mice, hamster and rat were used to study and evaluate plasmocin™ efficiency and compare it to commonly used antibiotics such as BM-cyclin, ciprofloxacin and mycoplasma removal agent (MRA). It was shown that mycoplasma infections were eradicated by plasmocin™, BM-cyclin, ciprofloxacin and MRA in 65%, 66.25%, 20%, and 31.25%, respectively, of infected cell cultures. However, re-infection with mycoplasmas after the period of 4 months occurred in 10–80% of the studied cell lines. Cell cytotoxicity and culture death was observed in 25, 17.5 and 10% of the treated cells, for plasmocin™, BM-cyclin and MRA, respectively. In this study, Plasmocin™ showed strong ability to eradicate mollicutes from our cell lines with minimal percentage of regrowth. However, due to its high cell cytotoxicity it should be used with caution especially when dealing with expensive or hard-to-obtain cell lines. Amongst the antibiotics tested, BM-cyclin was shown to remove mycoplasma with the highest efficiency.     

2194例男性泌尿生殖道支原体检测及药敏分析

目的研究男性泌尿生殖系统炎症患者解脲脲原体(Uu)和人型支原体(Mh)感染及对抗菌药物耐药情况,以了解本地区男性支原体的感染流行状况并指导临床合理选择使用抗菌药物。方法采用支原体鉴定药敏试剂盒,对男性生殖道感染者进行支原体培养及药敏试验。结果 2 194份标本中检出支原体636例,阳性率为28.96%,其中解脲脲原体阳性率为22.59%,人型支原体阳性率为0.96%,解脲脲原体和人型支原体混合感染阳性率为5.42%。支原体对抗菌药物药敏结果显示,抗菌活性较高的是多西环素、普拉霉素、四环素和交沙霉素。结论Uu是男性泌尿生殖系统支原体感染的主要病原体。临床治疗支原体感染时应尽量根据药敏结果选择敏感药物。普拉霉素、多西环素、四环素和交沙霉素可作为本地区非淋菌尿道炎经验用药的首选药物,避免使用喹诺酮类药物。     

重组酶聚合酶等温扩增快速检测肺炎支原体方法的建立和初步应用

摘要 目的：建立基于重组酶聚合酶扩增技术（RPA）技术快速检测肺炎支原体的方法。方法：本研究以肺炎支原体编码P1黏附蛋白为靶基因,利用Primer Premier 5软件进行引物、探针的设计,最终筛选出最佳引物。同时设计相应的实时荧光定量PCR（RT-PCR）引物用于后续的验证试验。对反应体系试剂比例、反应时间、反应温度、引物探针浓度进行确定。肺炎支原体、解脲支原体、人型支原体、肺炎克雷伯菌、肺炎双球菌、大肠杆菌和链球菌作为对照评估RPA检测肺炎支原体的特异性及敏感度。结果：RPA快速检测肺炎支原体方法仅需14 min,检测灵敏度达200 copies/mL;6种非肺炎支原体均不能扩增,特异性较高。结论：本研究建立了肺炎支原体的RPA快速检测方法,具有迅速、简便、经济等优势,为肺炎支原体的快速检测提供一个新的有利工具。     

一步法PCR检测大鼠肺支原体核酸的研究

一步法体外扩增结合Ｓｏｕｔｈｅｒｎ杂交检测Ｍ５３鼠肺支原体标准株,设计一对特异寡核苷酸引物及探针,合成、纯化、建立了特异、敏感、快速的检测手段。扩增产物经琼脂糖凝胶电泳鉴定,结果显示鼠肺支原体Ｍ５３株基因组ＤＮＡ７１０ｂｐ特异谱带。对５０只ＳＤ大鼠进行检测,结果ＰＣＲ方法检出率高于分离培养法,扩增产物行Ｓｏｕｔｈｅｒｎｂｌｏｔ杂交验证,采用碱性磷酸酶标记寡核苷酸探针,可与膜上特异靶ＤＮＡ序列杂交,而阴性对照无杂交信号。特异性实验检出１０ｐｇ的ＤＮＡ。充分说明一步法ＰＣＲ,具有高度、特异、灵敏、快速等优势,适应与大、小鼠监测中应用。     

肺炎支原体肺炎合并喘息患儿血清25（OH）D3水平与肺功能的相关性及其影响因素分析

摘要 目的：探讨肺炎支原体肺炎（MPP）患儿合并喘息的影响因素,分析血清25-羟维生素D3[25(OH)D-3]水平与MPP合并喘息患儿肺功能的关系。方法：选择2017年1月至2020年1月我院收治的90例MPP患儿,根据是否合并喘息将其分为MPP合并喘息组（39例）和MPP未合并喘息组（51例）。检测血清25(OH)D-3水平以及肺功能[最大呼气流速（PEF）、PEF占预计值百分比(PEF% pred)、第一秒用力呼吸容积（FEV₁）、用力肺活量（FVC）、FEV₁/FVC比值、FEV₁占预计值百分比（FEV1%pred）],Pearson相关性分析25(OH)D-3与MPP合并喘息患儿肺功能的关系,单因素及多因素Logistic回归分析影响MPP合并喘息的危险因素。结果：MPP合并喘息组血清25(OH)D-3水平、PEF、PEF% pred、FEV₁/FVC比值、FEV1%pred低于MPP未合并喘息组（P＜0.05）,Pearson相关性分析显示,MPP合并喘息组患儿血清25(OH)D-3水平与PEF、PEF% pred、FEV₁/FVC比值、FEV₁%pred均呈正相关（r=0.519、0.612、0.571、0.593,P＜0.05）。单因素分析显示,MPP合并喘息组年龄低于MPP未合并喘息组（P＜0.05）,病程长于MPP未合并喘息组（P＜0.05）,肺部啰音比例、嗜酸性粒细胞计数、MP-IgM抗体滴度高于MPP未合并喘息组（P＜0.05）。多因素Logistic回归分析结果显示低龄、肺部啰音、嗜酸性粒细胞计数增高、MP-IgM抗体滴度增加、25(OH)D-3减少是MPP合并喘息的危险因素（P＜0.05）。结论：MPP合并喘息患儿25(OH)D-3水平较低,低龄、肺部啰音、嗜酸性粒细胞计数增高、MP-IgM抗体滴度增加、25(OH)D-3缺乏为MPP合并喘息的危险因素,25(OH)D-3缺乏与MPP合并喘息患儿肺功能下降有关。     

穿透支原体LAMPs诱导NF-kB激活介导小鼠巨噬细胞凋亡

研究穿透支原体(Mpe)脂质相关膜蛋白(LAMPs)能否诱导小鼠巨噬细胞凋亡,并阐明其可能的分子机制,以了解Mpe潜在的致病性.用Annexin-V-FITC凋亡检测试剂盒和DNA Ladder方法检测Mpe LAMPs诱导体外培养的小鼠巨噬细胞系Raw264.7细胞的凋亡.以间接免疫荧光和Western blotting方法检测经Mpe LAMPs处理的小鼠巨噬细胞NF-κB的激活和NF-kB抑制剂吡咯啉烷二甲基硫脲(PDTC)对细胞凋亡的影响.结果表明:Mpe LAMPs能诱导小鼠巨噬细胞发生早期或晚期凋亡;Mpe LAMPs能诱导激活小鼠巨噬细胞的NF-κB,使其从细胞浆中转位到细胞核内;PDTC能显著地抑制经处理的小鼠巨噬细胞的NF-κB的激活,且能抑制Mpe LAMPs诱导的巨噬细胞发生凋亡.因此,Mpe LAMPs诱导小鼠巨噬细胞凋亡可能与NF-kB的激活有关,因而Mpe可能是一个重要的致病因素.     

不同剂量甲泼尼龙三联疗法对难治性肺炎支原体肺炎患儿免疫功能及炎性因子水平的影响

目的:探讨不同剂量甲泼尼龙三联疗法对难治性肺炎支原体肺炎患儿免疫功能及炎性因子水平的影响。方法:选取我院在2017年5月到2018年6月期间收治的120例肺炎支原体肺炎患儿,根据随机数字表法将其分为对照组、低剂量组、中剂量组和高剂量组,每组均为30例。对照组采用阿奇霉素联合头孢他啶进行治疗,低剂量组、中剂量组和高剂量组在对照组的基础上分别给予1 mg/(kg·d)、2 mg/(kg·d)、5 mg/(kg·d)的甲泼尼龙进行治疗。比较各组患儿的临床疗效、临床症状消失时间及不良反应发生情况,并比较各组患儿治疗前后CD3~+、CD4~+、CD8~+、红细胞沉降率(ESR)、C反应蛋白(CRP)、肿瘤坏子因子-α(TNF-α)、白介素-6(IL-6)水平。结果:高剂量组的总有效率高于对照组(P0.05)。高剂量组患儿临床各项症状消失时间短于中剂量组、低剂量组和对照组(P0.05),中剂量组和低剂量组患儿临床各项症状消失时间短于对照组(P0.05)。治疗后高剂量组、中剂量组、低剂量组的CD3~+、CD4~+水平高于对照组(P0.05)。治疗后高剂量组患儿的ESR、CRP、IL-6水平低于中剂量组、低剂量组和对照组,且中剂量组和低剂量组患儿的ESR、CRP、IL-6水平低于对照组(P0.05)。各组的不良反应发生率比较无统计学差异(P0.05)。结论:甲泼尼龙三联疗法治疗难治性肺炎支原体肺炎患儿疗效确切,可提高患儿免疫功能,且高剂量的甲泼尼龙三联疗法可促进患儿的临床症状改善,更明显地降低炎性因子的水平,安全可靠。     

Biochemical and genetic variation in Mycoplasma fermentans strains from cell line, human and animal sources

Aims:  To investigate the inter-strain variation in (i) substrate utilization and (ii) the restriction fragment length polymorphism (RFLP) pattern based on the distribution of an insertion element (IS 1550 ) in Mycoplasma fermentans strains, and to establish any correlation between subgroups within the species and their source or habitat.
Methods and Results:  Using a sensitive dynamic pH method, the pattern and kinetics of substrate utilization by a panel of 17 M. fermentans strains from various sources was determined. This study correlated the biochemical characteristics of these strains with RFLP patterns based on the distribution of an insertion sequence (IS 1550 ) with the sources of the strains. The test isolates were divided into four major groups according to the pattern of substrates metabolized. Interestingly, two strains isolated from cell lines in RFLP cluster I failed to utilize arginine. Ovine strains showed distinct substrate utilization patterns and produced RFLP patterns not previously encountered.
Conclusions:  All strains utilized glucose, but the ability to utilize arginine, fructose and N -acetyl glucosamine varied. There was also some correlation evident between the metabolic data and the RFLP clusters.
Significance and Impact of the Study:  This study has provided a better understanding of the biochemical and genetic diversity of M. fermentans strains from various sources.