Diagnostic Value Of Combined Carcino-embryonic Antigen Immunocytochemical Stain Examination And Laser Scanning Confocal Microscope In Patients With Meningeal Carcinomatosis

Objective: Epidemiological research indicated that Malignant Tumor Mortality ranked 1st among diseases of death in the world. So cancer is the increasingly serious diseases which threatens human's health and lives. And meningeal carcinomatosis is a special kind in the cancer. It is a kind of non-independent diseases and the tumor cells avert to the brain and spinal arachnoid and pia mater, but it could not discovered either grossly or microscopically in the parenchyma of central nervous system. In patients with solid tumor, the cancer rate was 5% and the mortality was high. And it infiltrate into cerebrocortex, which produce a serious of clinical syndrome. Its clinical symptoms were often earlier found to the primary disease and the clinical features have no special. Its earlier clinical symptoms only have signs of meningeal irritation and increased CSF pressure. Then the other symptoms in the nerves system have coming also. Symptom of the whole brain was the most serious yet. Of course, the signs of meningeal irritation are the most common clinical manifestation. MC also called carcinomatous meningitis because of it usually confused with meningitis. It is very difficult to diagnose in vivo by common imaging because it does not form the carcinoma mass. So many patients were delayed by these factors when they went to hospital. That endangered the burden of family and society seriously. But at present it changed by the method of cerebrospinal fluid cytology(CSFC), which is the primary examination method at home and abroad. May-Gruwald- Giemsa stain was the common cytologic method, but just observing cell morphological changes was hard in qualitative diagnosis when the tumor cell pleomorphism was untypical. In order to improve the accurate diagnosis rate in MC, our subject combined carcino-embryonic antigen immunocytochemical stain examination with double immunofluorescence staining by Laser Scanning Confocal Microscope (LSCM)——From cell morphological to molecular Level.Methods: The 42 patients with MC were Definitely diagnosed in our hospital. All patients CSF were collected in 24 to 48 hours, and produce cell smears from CSF by centrifugal precipitation. The CSF cell smears were used to be stain respectively,42 cases with May-Gruwald-Giemsa stain,29 cases with carcino-embryonic antigen Immunocytochemical stain and 17 cases with double immunofluorescence staining. There were 20 cases negative controls with carcino-embryonic antigen Immunocytochemical stain and double immuno- fluorescence staining respectively. Cell smears with the former two methods observed under light microscope . Cell smears with immunofluorescence staining observed by LSCM, and analyzed in locating, qualitative, quantitative and 3-D image reconstruction. According to the compareing analysis of sensitivity and specificity of three methods, concluded their superiority and inferiority.Results: Malignant cells were found in all of 42 cases for repeated CSF testing. The positive ratios with MC of May-Gruwald-Giemsa stain, carcino-embryonic antigen Immunocytochemical stain and double immunofluorescence staining were 85.71%(36/42), 79.31%(23/29) and 76.47% (13/17), in the first cerebrospinal fluid specimens respectively. Comparison of the positive rate of three methods have no statistical difference (P>0.05). The CEA positive cells with MC were red cytoplasm and membrane around blue nucleus under LSCM. The CEA negative cells both of patients with MC and negative controls just appeared blue nucleus. Quantitative Analysis of LSCM appeared that the average fluorescence intensity of negative controls about CEA and Nuclear DNA were (360.686-90.32) and (946.000-184.284) respectively. The CEA positive cells with MC were (2104.515-853.274) and (2710.135-793.863) respectively. When the MC group of CEA (+) compared with control group, the average fluorescence intensity both CEA and Nuclear DNA were significant differences (P<0.01); when the MC group of CEA(-) Compared with control group, the average fluorescence intensity of Nuclear DNA were significant differences (P<0.01), but the average fluorescence intensity of CEA were not statistical difference(P>0.05).Conclusions:1 May-Gruwald-Giemsa stain of CSFC is the main and basic method in diagnosing MC at present.2 Immunocytochemical stain of CSFC provide a powerful corroboration of qualitative diagnosis for MC.3 LSCM and immunofluorescence staining improved the diagnostic level of MC in locating, qualitative and quantitative. Especially quantitative analysis appeared that compared with control group, the average fluorescence intensity of Nuclear DNA of the MC group were polyploidy difference. It is valuable to diagnose MC when tumor markers in Immunocytochemical stain are negative.4 There are complementary diagnostic value of May-Gruwald- Giemsa stain, Immunocytochemical stain and Immuno- fluorescence staining.