CAJ | 학술논문

目的探讨免疫细胞化学染色及激光扫描共聚焦显微镜(LSCM)技术在脑膜癌病(meningeal carcinomatosis,MC)早期诊断中的应用价值.方法将所观察脑脊液标本分为MC组、非MC组(对照组),以Shandon Cytospin 4离心沉淀仪收集脑脊液细胞.分别进行Giemsa染色(42例)、联合癌胚抗原(CEA)免疫细胞化学染色(29例,对照20名)、免疫荧光双标记染色(17例,对照20名).CEA免疫细胞化学染色采用SP法,置于普通光学显微镜下观察;免疫荧光双标记染色以CyS荧光二抗及4,6-联脒-2-苯基吲哚分别标记抗CEA单抗及细胞核DNA,置于LSCM下观察,进行定位、定性、定量分析.结果 42例患者经脑脊液细胞学检查均发现异常细胞,首次脑脊液细胞学检查阳性率为85.7%(36/42),其中29例患者结合CEA免疫细胞化学染色,阳性率为79.3%(23/29),17例经免疫荧光双标记染色,13例阳性.MC组[CEA(+)者1694.69±478.06,CEA(-)者1543.04±364.71]与对照组(603.72±178.04)比较胞核DNA荧光相对含量[CEA(+)者t=21.386,CEA(-)者t=23.144,均P<0.01]差异有统计学意义;CEA荧光相对含量CEA(+)者(1407.04±275.30)与对照组(202.51±54.05)差异亦有统计学意义(t=42.934,P<0.01).结论免疫细胞化学染色是诊断MC的重要定性手段,结合LSCM可将MC的诊断提高到定性、定位与定量相结合的新水平.
목적 탐토면역세포화학염색급격광소묘공취초현미경(LSCM)기술재뇌막암병(meningeal carcinomatosis,MC)조기진단중적응용개치.방법 장소관찰뇌척액표본분위MC조、비MC조(대조조),이Shandon Cytospin 4리심침정의수집뇌척액세포.분별진행Giemsa염색(42례)、연합암배항원(CEA)면역세포화학염색(29례,대조20명)、면역형광쌍표기염색(17례,대조20명).CEA면역세포화학염색채용SP법,치우보통광학현미경하관찰;면역형광쌍표기염색이CyS형광이항급4,6-련미-2-분기신타분별표기항CEA단항급세포핵DNA,치우LSCM하관찰,진행정위、정성、정량분석.결과 42례환자경뇌척액세포학검사균발현이상세포,수차뇌척액세포학검사양성솔위85.7%(36/42),기중29례환자결합CEA면역세포화학염색,양성솔위79.3%(23/29),17례경면역형광쌍표기염색,13례양성.MC조[CEA(+)자1694.69±478.06,CEA(-)자1543.04±364.71]여대조조(603.72±178.04)비교포핵DNA형광상대함량[CEA(+)자t=21.386,CEA(-)자t=23.144,균P<0.01]차이유통계학의의;CEA형광상대함량CEA(+)자(1407.04±275.30)여대조조(202.51±54.05)차이역유통계학의의(t=42.934,P<0.01).결론 면역세포화학염색시진단MC적중요정성수단,결합LSCM가장MC적진단제고도정성、정위여정량상결합적신수평.
Objective To explore the early diagnostic value of carcino-embryonic antigen (CEA) immunocytochemistry examination combined with laser scanning confocal microscope (LSCM) in the patients with meningeal carcinomatosis (MC).Methods The patients were divided into experimental group (patients with MC) and control group (patients without MC).Thermo electron corporation shandon cytospin 4 centrifuge was used in the cytologic examination of cerebrospinal fluid (CSF),whose function was to produce a monolayer of cells onto a glass slide from CSF.Giemsa staining was used in 42 cases.The CEA immunocytochemistry staining was used in 29 cases and 20 controls.The double immunofluorescence staining was used in 17 cases and 20 controls.SP staining method was used in the CEA and the results were observed under the light microscope.Nuclear DNA and CEA were stained with fluorescent probe DAPI and Cy5 respectively and the results of double immunofluorescenee staining were observed by the laser scanning confocal microscope.Results There was a high positive rate in cytologic examination of CSF,and malignant cells were found in all of 42 cases for repeated CSF testing.The positive rate of routine CSF cytologic examination and CEA immunecytochemistry examination was 85.7% (36/42) and 79.3% (23/29) respectively in the first CSF specimens.There were 17 cases using double immunofluorescence staining and observed by LSCM,and the positive rate was 13/17.Compared with experimental group and control group,fluorescent value both nuclear DNA (CEA(+) 1694.04±478.06,CEA(-)1543.04±364.71,control group 603.72±178.04,t=21.386,23.144,both P<0.01) and CEA (CEA(+)1407.04±275.30,control group 202.51±54.05,t=42.934,P<0.01) were significantly different.Conclusion Immunocytochemistry examination of CSF is an important early qualitative diagnosis method for MC.LSCM improved the level of locating,qualitative and quantitative diagnosis of MC.