To Establish A Platform For Diagnosing The Niemann-Pick Disease Type C

Part one Filipin staining of cultured skin fibroblasts in Diagnosing Niemann-Pick disease type CObjective The diagnostic significance of filipin staining of cultured skin fibroblasts in diagnosing Niemann-Pick disease type C has been evaluated.Methods After fibroblasts of the mouse skin were cultured with lipoprotein- deficient fetal bovine serum(LPDS)for 5 days, and then co-cultured with human low density lipoprotein (LDL) for 24 hours, the cultured skin fibroblasts were subjected to filipin staining. Under a confocal microscope, photography was harvested with excitation of violet light. At the same time, both mutant and wild-type mice were analyzed and evaluated genetically and neuropathologically.Results Filipin staining demonstrates an intense punctate pattern of fluorescence concentrated around the nuclei of the cultured cells, consistent with the aberrant accumulation of the unesterified cholesterols in the npc mice. On the contrary, the filipin fluorescence was faintly distributed inside the cytoplasma.Conclusion Filipin stain can diagnose the Niemann-Pick disease type C.Part two One Case report of a typical patient of Nimman-Pick disease type CObjective One typical patient of the Niemann-Pick disease type C on the clinic ground, biochemical examination and gene analysis was identified. Through this case, we have successfully established the system methlods to idenfity NPC in China. Methods To analysis the materials of the typical patient of the NPC, filipin stain of the cultured skin fibroblast,and sequence analysis of the genealogy in order to definitively diagnose the NPC.Results The patient have a typical clinic presentation. Filipin staining demonstrates an intense punctate pattern of fluorescence concentrated around the nucleus, consistent with the accumulation of unesterified cholesterol. Gene sequence analysis show that there are two mutation points, 1142 ( M1142T ) and 1186 ( R1186H ) respectively, coming from mother and father. The patient can be definitively diagnosed as NPC.Conclusion Skin fibroblast filipin stain and gene sequence analysis can final diagnosis the Nimann-Pick disease type C.