肺炎支原体肺炎患儿血清SP-D、Gal-3、CCL5水平与炎症因子和预后不良的关系研究

摘要 目的：探讨肺炎支原体肺炎（MPP）患儿血清表面活性蛋白D（SP-D）、半乳糖凝集素-3（Gal-3）、C-C基序趋化因子配体5（CCL5）水平与炎症因子和预后不良的关系。方法：选取2020年1月～2021年1月我院收治的165例MPP患儿为MPP组,另选取54例健康儿童为对照组。采用ELISA法检测血清SP-D、Gal-3、CCL5、白细胞介素-6（IL-6）、IL-8水平,放射免疫法检测血清肿瘤坏死因子-α（TNF-α）水平。采用Pearson/Spearman相关系数分析MPP患儿血清SP-D、Gal-3、CCL5水平与IL-6、IL-8、TNF-α水平的相关性。通过多因素Logistic回归分析MPP患儿预后不良的影响因素。采用接受者操作特征（ROC）曲线分析血清SP-D、Gal-3、CCL5水平对MPP患儿预后不良的预测价值。结果：与对照组比较,MPP组患儿的血清SP-D、Gal-3、CCL5以及IL-6、IL-8、TNF-α水平升高（P＜0.05）。MPP患儿的血清SP-D、Gal-3、CCL5水平与IL-6、IL-8、TNF-α水平均呈正相关（P＜0.05）。血清SP-D、Gal-3、CCL5、IL-6、TNF-α水平较高、病变类型为大片状阴影、热程较长是MPP患儿预后不良的危险因素（P＜0.05）。血清SP-D、Gal-3、CCL5三项联合预测MPP患儿预后不良的曲线下面积（AUC）为0.926,明显大于三项指标单独预测的0.842、0.794、0.771。结论：MPP患儿血清SP-D、Gal-3、CCL5水平升高,与炎症因子和预后不良密切相关,可作为MPP患儿预后不良的预测指标。     

甘草酸对巨噬细胞抗绵羊肺炎支原体感染的调控作用研究

为观察甘草酸在小鼠巨噬细胞系RAW264.7抗绵羊肺炎支原体(Mycoplasma ovipneumoniae,MO)感染中的作用,实验利用CCK-8细胞活性检测找到最佳甘草酸处理浓度;检测甘草酸对受MO感染的巨噬细胞活性的影响。流式细胞仪检测甘草酸对RAW264.7巨噬细胞生长周期的影响;ELISA检测甘草酸对受MO感染的巨噬细胞分泌TNF-α的影响;Western blot检测细胞凋亡因子Bax、Bad的表达情况。RT-PCR检测凋亡和自噬相关基因的表达情况。结果显示,浓度为12μmol/L的甘草酸显著升高RAW264.7的活性(P=0.012 9),且处于G1期的细胞数减少,G2期的细胞数增加。甘草酸(12μmol/L)可提高受MO感染的RAW264.7的增殖率(P=0.034 0),培养上清中TNF-α含量升高(P=0.015 2),巨噬细胞中促凋亡蛋白Bax表达量增加,但基因caspase 3和caspase 9的表达量显著下调(P<0.000 1),自噬相关基因Atg 7和Beclin 1表达量显著升高(P<0.000 1)。结果提示在MO感染巨噬细胞引起免疫抑制的情况下,甘草酸可通过促增殖、抑凋亡、促进TNF-α的表达、增加自噬来起到免疫调控作用。     

肺炎支原体分型与肺炎患儿临床症状相关性

目的研究肺炎支原体分型与肺炎患儿临床症状的相关性。方法选择2017年11月至2018年5月在郑州大学附属儿童医院儿科治疗的确诊为肺炎支原体(MP)肺炎的患儿86例。采用自行设计的调查表研究患儿基本资料,包括性别、年龄、既往史、入院24h临床症状、实验室指标、影像学症状等。利用实时定量PCR技术分析样本中肺炎支原体基因拷贝数,并设计引物分析MP基因分型。结果共检出MP低拷贝量患儿15例(17.44%),中拷贝量患儿24例(27.91%),高拷贝量患儿47例(54.65%);P1-I型MP 80例(93.02%),P1-II型MP 6例(6.98%)。患儿MP基因拷贝数与病程≥10d,体温39℃,畏寒或寒战,GRA%,GRA,CRP,PCT,大片状实变/不张,胸腔积液呈正相关,且为弱或中等相关。MP基因拷贝数与LYM%,LYM呈负相关,且为弱相关。患儿MP分型与体温39℃呈负相关,且为弱相关。结论 MP肺炎患儿上呼吸道MP数量与患儿临床表现,实验室指标和影像学特征存在一定相关性,因此上呼吸道MP数量可成为MP肺炎病情评估的指标之一。     

常见支原体感染误诊与对策

临床支原体感染易发生误诊,不但加重患者精神和经济负担,而且延误患者病情,甚至留下后遗症。为此,我们综述了常见支原体感染误诊的特点和原因,深挖其中思维和思想根源,探讨减少或避免该类误诊现象的策略,旨在为临床医护人员和检验技术人员提供参考建议。     

Chlamydia and mycoplasma infections during pregnancy and their relationships to orofacial cleft

Serum antibodies to Mycoplasma pneumoniae and Chlamydia trachomatis have been studied in a group of newborns with orofacial cleft (OC) and their mothers (n = 59) as compared to a control group of healthy newborns and their mothers (n = 40) assayed by ELISA and Western blot analysis. In the first group, IgG antibodies to M. pneumoniae were found by ELISA in 12 newborns with OC and 22 mothers, while IgA antibodies were detected only in 5 and 11 cases, respectively. IgM antibodies indicating an acute infection were found in 2 mothers only. IgG antibodies to C. trachomatis were found in 2 newborns with OC and 4 mothers. In the control group, IgG antibodies to M. pneumoniae were found in 3 newborns and 7 mothers. IgG antibodies to C trachomatis were observed in 1 newborn and 1 mother, while IgM antibodies to C trachomatis were present in 1 mother only. Immunoblot analysis revealed in newborns with OC and their mothers C. trachomatis-specific bands associated with MOMP 1, 29 kDa, 45 kDa, and heat shock proteins (HSP) 60 and 70. Based on these results we suggest that the risk associated with the exposure to M. pneumoniae and/or C. trachomatis is so far unknown and further study is needed for its elucidation.     

Target selection and deselection at the Berkeley Structural Genomics Center

At the Berkeley Structural Genomics Center (BSGC), our goal is to obtain a near-complete structural complement of proteins in the minimal organisms Mycoplasma genitalium and M. pneumoniae, two closely related pathogens. Current targets for structure determination have been selected in six major stages, starting with those predicted to be most tractable to high throughput study and likely to yield new structural information. We report on the process used to select these proteins, as well as our target deselection procedure. Target deselection reduces experimental effort by eliminating targets similar to those recently solved by the structural biology community or other centers. We measure the impact of the 69 structures solved at the BSGC as of July 2004 on structure prediction coverage of the M. pneumoniae and M. genitalium proteomes. The number of Mycoplasma proteins for which the fold could first be reliably assigned based on structures solved at the BSGC (24 M. pneumoniae and 21 M. genitalium) is approximately 25% of the total resulting from work at all structural genomics centers and the worldwide structural biology community (94 M. pneumoniae and 86 M. genitalium) during the same period. As the number of structures contributed by the BSGC during that period is less than 1% of the total worldwide output, the benefits of a focused target selection strategy are apparent. If the structures of all current targets were solved, the percentage of M. pneumoniae proteins for which folds could be reliably assigned would increase from approximately 57% (391 of 687) at present to around 80% (550 of 687), and the percentage of the proteome that could be accurately modeled would increase from around 37% (254 of 687) to about 64% (438 of 687). In M. genitalium, the percentage of the proteome that could be structurally annotated based on structures of our remaining targets would rise from 72% (348 of 486) to around 76% (371 of 486), with the percentage of accurately modeled proteins would rise from 50% (243 of 486) to 58% (283 of 486). Sequences and data on experimental progress on our targets are available in the public databases TargetDB and PEPCdb.     

建立一种反向斑点杂交法检测肺炎支原体23S rRNAV区A2063G基因突变的方法

目的建立检测肺炎支原体（Mycoplasma pneumoniae,MP）23S rRNA V区2063基因型的反向斑点杂交方法,并与PCR产物直接测序法的结果进行比较分析。方法19例自临床咽拭子标本分离培养的MP,用自行设计的反向斑点杂交法检测23S rRNA V区2063基因型,同时对相应序列测序分析。抽提标准菌株FH和127份经PCR测定MP阳性的临床标本基因组DNA,用MP阴性的基因组DNA抽提物5份作阴性对照,用反向斑点杂交法检测23S rRNA V区2063基因型。结果19例分离培养的MP,经反向斑点杂交法检测15株有23S rRNA V区基因A2063G位点突变;4株为2063A,与测序结果一致（Kappa一致性检验,P〉0.05）。经反向斑点杂交法检测,127份MP阳性的基因组DNA抽提物中有122份标本为A2063G位点突变,标准菌株FH和2份DNA抽提物的2063位点碱基为A,还有3份抽提物标本的2063位点碱基既有A又有G,5份阴性对照均无显色。结论反向斑点杂交方法能快速、准确检测临床MP 23S rRNA V区2063基因型。     

In vivo virulence of Mycoplasma hyopneumoniae isolates does not correlate with in vitro adhesion assessed by a microtitre plate adherence assay

Aims:  Adherence of Mycoplasma hyopneumoniae to the ciliated epithelial cells of the porcine respiratory tract is considered an important first step in the pathogenesis of enzootic pneumonia. It was the aim of this study to verify the usefulness of in vitro adhesion as a virulence marker.
Methods and Results:  Adherence capacity to immobilized cilia from porcine tracheal epithelial cells of three low, two moderately and two highly virulent M. hyopneumoniae field isolates was determined by a microtitre plate adherence assay.
Conclusions:  No significant differences between the isolates were demonstrated.
Significance and Impact of the Study:  The results suggest that mechanisms other than adherence might be responsible for the observed differences in virulence of these field isolates or that the in vitro assay does not adequately reproduce in vivo adherence conditions.     

非淋菌性泌尿生殖道炎支原体感染及耐药变迁

目的探讨解脲支原体（Uu）及人型支原体（Mh）在非淋菌性泌尿生殖道炎的感染状况,分析近3年来支原体的感染及耐药变迁。方法采用培养法（Mycoplasma IST）对NGGU的分泌物进行支原体的培养、体外药敏试验及分析。结果支原体阳性率为47．0％,其中Uu阳性率为34．0％,Uu＋Mh阳性率为11．4％,Mh阳性率为1．6％;交沙霉素、强力霉素等药物的敏感率无明显变化,而喹诺酮类的耐药率则呈逐渐上升趋势,环丙沙星及氧氟沙星的耐药率分别从2002年的38．9％及40．0％上升到了2004年的79．2％及66．4％。结论支原体的耐药率正逐步增加,其中喹诺酮类的高耐药率提示其在治疗支原体感染中的地位改变,泌尿生殖道支原体的药物敏感监测对临床治疗具有重要意义。