构建玻片蛋白质芯片的实验研究

目的:探讨玻片蛋白质芯片的构建方法及其中间操作条件的选择与优化。方法:利用生物芯片点样仪将超微量蛋白质点到经特殊处理的玻璃片表面,然后选用不同的化学试剂对玻片背景进行封闭;再用荧光素标记的蛋白质与点样蛋白杂交;最后用生物芯片分析仪扫描成像并进行分析,比较不同条件下芯片的显示效果。结果:蛋白质样品与片基稳定结合,并保持原活性状态;封闭试剂选用酪蛋白或明胶效果较佳;点样探针与其特异蛋白质抗体可稳定结合,结合效果与点样蛋白浓度在一定范围内呈正相关;在1.6 cm×1.6 cm的玻璃片面积上,构建了36×36=1269点的蛋白质芯片。结论:本实验条件下,蛋白质抗原或抗体可以稳定地固定于经过处理的玻璃片表面.制成免疫型蛋白芯片,并可通过随后的抗原抗体反应与荧光素标记的相应抗体或抗原结合,用生物芯片分析仪可对其荧光信号进行检测分析。     

Clarke's column in sporadic amyotrophic lateral sclerosis

Summary Histological, ultrastructural and morphometrical observations on Clarke's column were carried out in 18 patients with sporadic amyotrophic lateral sclerosis (ALS) and 15 age-matched control subjects. Of the 18 ALS patients 6 had been on a respirator before death. Bunina bodies were found in the neuronal cytoplasm in 7 of the 12 non-respirator-supported ALS patients and in 3 of the 6 respirator-supported patients. The number of spheroids was significantly higher in the non-respirator-supported patients (P<0.01) than in the control subjects; however, the number in the respirator-supported patients was about equal to that in the controls. The number of neurons in Clarke's column in the non-respirator-supported ALS patients was not reduced, but in the respirator-supported patients they tended to disappear with time after respiratory support. These findings suggest that Clarke's column neurons are also involved primarily in the disease process in sporadic ALS. However, they may begin to disappear only after the patients require respiratory support.Supported in part by a research grant for CNS degenerative diseases from the Ministry of Health and Welfare, Japan     

Pretransplant Exposure to Donor HLA-DR Antigen in Random Transfusion Units and the Development of Donor Antigen-Specific Hyporeactivity

Our previous studies have shown that the in vitro assay of donor antigen-specific hyporeactivity is a useful marker for identifying solid organ transplant recipients (kidney, lung and heart) at low risk for immunologic complications (i.e., late acute rejection episodes and chronic rejection). Donor antigen-specific hyporeactivity is defined as a significantly decreased post- vs. pretransplant proliferative response to donor antigens while response to third-party controls remains unchanged. We analyzed whether exposure to the same HLA-DR antigen pretransplant via random blood transfusion and posttransplant via the transplanted organ influenced the development of hyporeactivity. Thirty previously nontransfused recipients, each receiving two 150 ml pretransplant random blood transfusions, were assessed for hyporeactivity at 1 year posttransplant. Of the 12 recipients with pretransplant exposure to kidney HLA-DR via transfusions, 6 (50%) developed hyporesponsiveness; in contrast, of the 18 recipients who were not preexposed, only 3 (15%) exhibited this form of immunomodulation. Of interest, 2 of the 3 hyporesponsive recipients who were not preexposed, received units containing HLA-DR antigens previously shown to share crossreactive epitopes with the kidney HLA-DR. In conclusion, these results suggest a increased incidence in the development of hyporeactivity in patients receiving pretransplant transfusions which share an HLA-DR antigen with the transplanted kidney.     

Hemorrhagic fever with renal syndrome virus infection in livers studied by in situ hybridization and immunohistochemistry

ＨｅｍｏｒｒｈａｇｉｃｆｅｖｅｒｗｉｔｈｒｅｎａｌｓｙｎｄｒｏｍｅｖｉｒｕｓｉｎｆｅｃｔｉｏｎｉｎｌｉｖｅｒｓｓｔｕｄｉｅｄｂｙｉｎｓｉｔｕｈｙｂｒｉｄｉｚａｔｉｏｎａｎｄｉｍｍｕｎｏｈｉｓｔｏｃｈｅｍｉｓｔｒｙＹａｎｇＳｈｏｕｊｉｎｇ（杨守京）；Ｌ...     

大鼠胃黏膜癌变过程中细胞增殖与端粒酶活性关系

目的　探讨大鼠胃黏膜癌变发生发展过程中DNA含量、增殖细胞核抗原 (proliferatingcellnuclearantigen ,PCNA)及端粒酶活性两者之间的关系。方法　采用甲基硝基亚硝基胍 (MNNG)、10 %NaCl混合液及 40 %乙醇灌胃建立大鼠胃癌模型 ,设立正常组、模型组 ,分别采用免疫组化、流式细胞仪及端粒重复扩增微孔板杂交法检测大鼠胃黏膜PCNA、DNA含量及端粒酶活性。结果　本造模方法可诱发出浅表性胃炎 (chronicsuperficialgastritis,CSG)、萎缩性胃炎 (chronicatrophicgastritis,CAG)、肠化生 (intestinalmetaplasia,IM)、异型增生 (dysplasia,Dys)、胃腺癌(gastricadenocarcinoma,GC)等病变。在大鼠癌变过程中 ,随着胃黏膜癌变的发展 ,DNA含量、增殖细胞核抗原表达及端粒酶活性均呈递增趋势 ,并且两者与端粒酶活性具有高度的相关性 (P <0 .0 1,P <0 .0 1)。结论　大鼠胃黏膜癌变发生发展过程中 ,端粒酶的激活与胃黏膜组织细胞的恶性增殖具有重要的相关性。     

血清前－S_2蛋白检测的临床意义

采用ＥＬＩＳＡ法检测１０４例各类型乙型肝炎（乙肝）患者的血清前－Ｓ２抗原（Ｐｒｅ－Ｓ２Ａｇ）及其抗体（Ｐｒｅ－Ｓ２Ａｂ），结果表明，Ｐｒｅ－Ｓ２Ａｇ的出现与ＨＢｓＡｇ、ＨＢｅＡｇ、ＨＢｃＡｂ呈显著相关性（Ｐ值均＜０．００５），主要见于乙肝急性期及慢性乙肝患者，说明病毒复制活跃、传染性强。而Ｐｒｅ－Ｓ２Ａｂ阳性仅见于急性乙肝恢复期。     

子宫内膜异位症患者子宫内膜人白细胞抗原的免疫组化研究

目的 研究子宫内膜异位症(内异症)患者在位和异位子宫内膜人白细胞抗原HLA—DR的表达。方法 采用酶标链霉亲合素—生物素(LSAB)技术对19例内异症不孕患者在位和异位子宫内膜腺细胞HLA—DR抗原的表达进行免疫组化研究，以19例非子宫内膜异位症的不孕患者作为对照。结果 内异症患者在位和异位子宫内膜腺细胞HLA—DR抗原的表达较对照组明显增强(P＜0．01)。结论 子宫内膜HLA—DR抗原的异常表达可能参与了内异症免疫学异常的发生。     

癌胚抗原的测定及意义

癌胚抗原（ｃａｒｃｉｎｏｅｍｂｒｙｏｎｉｃａｎｔｉｇｅｎ，ＣＥＡ）作为肿瘤标志物之一，近年在肿瘤，特别是恶性肿瘤的诊断、监测和判断预后等方面得到了广泛地应用，是较灵敏和可靠的指标．本文就ＣＥＡ的测定、意义及不同标本测定的差别等作一综述     

Haemangiopericytomas: the prognostic value of immunohistochemical staining with a monoclonal antibody to proliferating cell nuclear antigen (PCNA)

Forty-two cases of haemangiopericytoma were studied retrospectively using immunohistochemical staining with PC10, a monoclonal antibody to PCNA. The percentage of tumour cells with positive staining for PCNA was found to correlate well with histological grading. Clinical follow-up data were available in 25 adults and showed no known deaths in 11 cases with a low proportion (less than 14%) of positive cells. Out of 14 cases with a high number (greater than or equal to 14%) of positive cells, seven patients are known to have died, two had metastases, and in a further two there have been multiple recurrences of tumour. DNA flow cytometry was performed on 26 cases but this showed no correlation with PC10 staining or clinical outcome. Staining with PC10 may be of particular value in the identification of patients at greatest risk of rapid tumour metastasis and early death.     

前列腺癌患者PSA、PSAD、PSAT测定

目的：探讨前列腺特异抗原（PSA）及前列腺特异抗原密度（PSAD）、前列腺特异抗原移行区密度（PSAT）作为前列腺癌肿瘤标志物的临床价值。方法：选择前列腺癌患者30例、前列腺增生患者30例、健康志愿者20例，采用放射免疫法检测PSA并计算PSAD、PSAT，比较3组间的变化情况。结果：前列腺增生组PSA升高，与对照组相比差异有统计学意义（P＜0．05），前列腺癌组PSA、PSAD、PSAT均升高，与对照组及前列腺增生组对比差异有统计学意义（P＜0．001）。结论：PSA作为前列腺癌肿瘤标志物有重要临床价值，但单纯测定血PSA含量有时不易区分前列腺增生与前列腺癌；PSAD、PSAT在前列腺增生与前列腺癌的鉴别诊断中优于PSA。