骨髓基质细胞固紫B盐染色法的建立及其临床意义

Fast violet B salt staining for bone marrow stromal cells and its clinical significance

目的建立骨髓基质细胞（BMSC）固紫B盐细胞化学染色法，并评估其临床应用价值。方法将传代培养的BMSC、骨髓、胸腔积液及腹腔积液标本制成涂片，行固紫B盐染色，观察细胞的着色情况及阳性BMSC形态特征，计数和比较再生障碍性贫血组（AA）、增生性贫血组与健康对照组骨髓标本的BMSC数；并以健康对照组BMSC数的下限值作为AA的诊断界点，随机双盲法评估该方法的诊断价值，计算敏感度、特异度、阳性似然比及阴性似然比。结果BMSC胞质染成紫红色，胞核不着色，其他细胞（粒、红、巨三系，单核、巨噬细胞，淋巴细胞，浆细胞，间皮细胞等）均不染色；从组BMSC计数为（1．07±0．29）个，增生性贫血组为（2．26±0．37）个，健康对照组为（1．58±0．33）个，从组BMSC计数明显低于增生性贫血组和健康对照组（q值分别为24．29、8．83，P均〈0．01），健康对照组低于增生性贫血组（q=16．41，P〈0．01）；本法诊断AA的敏感度、特异度、阳性似然比、阴性似然比分别为90％、93％、12．86、0．11。结论固紫B盐染色方法操作简便，特异性强，对骨髓的增生程度评价及贫血的鉴别诊断具有较高的诊断价值。

Objective To establish a quick method to identify BMSC by fast violet B salt staining and evaluate the clinic value. Methods Smears of separated and cultured BMSC, bone marrow, pleural and ascitic fluids were made, then the staining of fast violet B salt was performed. The BMSC in aplastic anemia （AA）, high hyperplasia and normal groups were counted and compared with each other. Meanwhile, the diagnostic value of this method to AA was evaluated. Results The cytoplasm of BMSC presented mauve, while the nucleus were negative, other cells such as myelocytes, nucleated erythrocytes, megakaryocytes, monocytes, macrophages, lymphocytes and plasmacytes were negative. The count of BMSC in AA, high hyperplasia and normal group was 1.07 ±0.29, 2. 26±0.37 and 1.58±0.33, respectively. Significant differences were found between AA and high hyperplasia groups, AA and normal groups, high hyperplasia and normal groups, respectively （P〈0.01）. The sensitivity, specificity, positive likelihood ratio and negative likelihood ratio of this method for diagnosis of AA were 90%, 93%, 12. 86 and 0.11, respectively. Conclusions The fast violet B salt staining is simple and convenient. It could be used to identify BMSC and play an important role in judging the hyperplasia extent and differentiation of AA.