正常人和肝病患者血中“致密体”及其与纳米细菌的关系

Characteristics of "dense bodies" in blood of patients with chronic hepatopathy or healthy people and their correlation with nanobacteria

目的研究正常和肝病患者血液中圆球体样微生物的生物学特性,初步确定纳米细菌与该种可滤过物的关系。方法将正常人和肝病患者培养阴性的血培养物进行细菌L型、真菌和厌氧菌培养。同时取沉淀用透射电镜观察。L型培养液用0.45μm和0.22μm滤器过滤,接种RPMI1640培养基,细胞培养条件下培养45d,用鼠抗纳米细菌单克隆抗体8D10免疫组化和钙盐染色进行纳米细菌鉴定。血浆纳米细菌培养阳性者12000×g离心后,进行普通和L型细菌培养。结果36/39患者和60/60健康对照血培养液中呈现类似于L型的巨型体、圆球体、原生小体的不明微生物。电镜观察圆球体内为电子致密样物质,周围未见细胞壁结构。电镜和光镜下可见其粘附在红细胞上或存在于红细胞内。滤过后培养物钙盐染色有5/39阳性,但纳米细菌免疫组化染色均阴性。纳米细菌阳性的培养物转种,未见一般细菌、细菌L型和上述血中"致密体"。结论血培养中致密体样微生物与纳米细菌无关,可能在维持机体正常免疫功能上具有一定意义。

Objectives To study the characteristics of spheroplast-like organisms in blood of chronic hepatopathy and healthy people,and discuss the relationship between nanobacteria（NB） and these novel filterable organisms. Methods Common blood cultures of 39 and 60 cases of hepatopathy or normal control were sub-cultivated respectively in anaerobic circumstance or with L-forms media,and observed with Transmission Electron Microscopy（TEM）. After 7 days,filtrates of them were subcuhured in RPMI 1640 media,and incubated in cell culture condition for 45 days. Then identified by immunohistochemistry stain（IHC） with anti-nanobacteria antibody 8D10, and calcific stain. Furthermore, pellets of 1640 cultures with NB positive were inoculated to SA or L-form liquid medium after 12 000 × g centrifugation. Results Novel organisms in common blood cultures were appeared for diseased（36/39） and normal humans（60/60） , resembled large body, sphero- plast, and elementary body of L-forms. Some of them adhering erethrocytes, Calcific stain of Filtered human blood cultures （ membrane filters with a pore size of 0.22 μm, 0.45 μm） were 5/39 positive and immunohistochemistry stains were all negative. Large and small ＂dense bodies＂ without cell-wall were observed with TEM. Ordinary bacteria,novel organisms, large bodies,spheroplasts, and elementary bodies of L-forms were not observed when cultures with NB were sub-cultivated to common bacterial media and L-forms media. Conclusions Novel organisms in blood are not related to nanobacteria. They possibly possess an important role in keeping normal immune function.