Immunobead and Density Gradient Purification of Paget Cells: In vitro Studies of Proliferation

Previous studies using primary monolayer cultures of epithelial cells from the involved epidermis of patients with mammary and extramammary Paget's disease investigated whether Paget cells proliferate as other malignant cells do. Although epithelial monolayers from the involved skin were maintained for approximately 45 days, no permanent cell lines were established. The proportion of carcinoembryonic antigen (CEA)-positive cells did not increase in the long-term cultures. Herein, we report studies of whether there is a real reduction of Paget cell numbers or if this is merely a decrease in the expression of CEA by the cells. Furthermore, we investigated whether Paget cells survive longer when cultured free from any potential inhibitory keratinocytes or other epidermal cells. Skin samples were obtained from one patient with mammary Paget's disease and three with extramammary Paget's disease; epidermal cells were cultured in vitro. An enrichment of Paget cells was carried out from the cultured epidermal cells by combining an anti-epithelial membrane antigen monoclonal antibody, binding to immunobeads, and density gradient centrifugation in Nycodenz. The separated cells were re-cultured in Keratinocyte-SFM serum-free media. The proportion of CEA-positive cells did not increase in the cultures, and the purified cells did not show any increase in survival times compared to the non-purified cultured cells. These results suggest that the decrease of CEA-positive cells noted during culture results from a decline in expression of CEA in the Paget cells. Paget cells in the involved epidermis do not proliferate significantly and thus differ from many other malignant cells.     

急性白血病非系限性分化抗原的动态研究

为探讨仅表达非系限性分化抗原的急性白血病的细胞起源，采用单克隆抗体免疫酶标和聚合酶链反应技术分析了１２例初诊时仅表达非系限性分化抗原的急性白血病化疗后免疫表型及免疫球蛋白重链（ＩｇＨ）和Ｔ细胞受体（ＴＣＲ）γ基因重排的变化。结果表明：初诊时仅表达ＣＤ３８抗原的５例急性白血病，化疗后３例出现Ｔ细胞相关抗原表达，并伴ＴＣＲγ基因重排；５例初诊时仅表达ＨＬＡ－ＤＲ或ＣＤ９的急性白血病，化疗后４例出现Ｂ细胞相关抗原表达，均伴有ＩｇＨ基因重排；２例无任何抗原表达者，化疗后１例表达Ｂ细胞相关抗原，另１例表达骨髓细胞相关抗原。提示免疫标志的动态研究，有助于初诊时免疫学无法分类急性白血病细胞起源的确定。     

Variation of prostate-specific antigen expression in different tumour growth patterns present in prostatectomy specimens

Summary A series of 55 randomly chosen radical prostatectomy specimens was analyzed for expression of prostate-specific antigen (PSA) by immunohistochemical techniques. Tissue sections were selected in such a manner that in addition to glandular benign prostatic hyperplasia (BPH), one or more different prostatic tumour growth patterns were present. Four monoclonal antibodies, directed against three different PSA epitopes, and one polyclonal anti-PSA antiserum were used. Expression of PSA was compared with that of prostate-specific acid phosphatase (PAP), recognized by two different polyclonal antisera. A critical dilution aimed at a maximum of staining intensity on BPH tissue sections was chosen for all antibodies. Anti-PSA and anti-PAP antisera stained essentially all BPH samples (over 90%). Irrespective of the nature of the antibodies used, PSA expression was found to be decreased in prostatic carcinoma. A clear cut relationship was found between immunoreactivity for PSA and the degree of differentiation of the tumour area. Under the experimental conditions used the PSA monoclonal antibodies stained only 1 out of 10 undifferentiated carcinomas, whereas 50% to 70% of the well- and moderately-differentiated carcinomas showed immunoreactivity. This correlation was less pronounced with the PAP staining pattern. If the PSA antibody titer was raised the percentage of clearly staining undifferentiated carcinomas could be considerably increased (up to 60%–100%), indicating that PSA expression is not absent, but lowered in most (if not all) undifferentiated carcinomas.     

PCNA在乳腺病变组织中的表达与病理学特性关系

本文运用增殖细胞核抗原(PCNA)单克隆抗体对88例乳腺病变组织进行免疫组化研究,并与乳腺癌组织学分级和淋巴结转移作了对比分析,结果显示:乳腺良性病变阳性率为16%,乳腺癌为73.02%,两者比较有显著性差异P<0.01),pCNA指数与乳腺癌组织学分级淋巴结转移密切相关.作者认为PCNA的表达对区分良恶性病变及组织学分级,淋巴(?)转移(?)为客观可靠,是(?)癌预后判断的良好指标.     

单克隆抗体对溶组织内阿米巴吞噬红细胞的影响

用具有抗细胞膜活性和抗细胞核、胞浆活性的单克隆抗体３Ｆ７和４Ｇ６与致病性溶组织内阿米巴ＨＭ－１∶ＩＭＳＳ株滋养体３７℃孵育，在孵育即刻、５ｍｉｎ、１０ｍｉｎ加入红细胞，检测滋养体吞噬红细胞的能力，并以正常小鼠血清和抗克氏锥虫抗体ＴＣＥ０４作为对照。结果，经单克隆抗体３Ｆ７孵育即刻的ＨＭ－１∶ＩＭＳＳ虫株滋养体吞噬红细胞能力明显减弱（Ｐ＜０．００１），而４Ｇ６抑制滋养体吞噬红细胞的作用不如前者明显。提示，抗细胞膜单克隆抗体对抑制滋养体吞噬红细胞有显著作用，但随着时间的推移，抑制作用明显减弱。     

金地鼠颊囊癌模型PCNA,BrdU及光镜观察结果的相关分析

癌变的早期诊断对提高疗效具有特殊意义，为了寻求简捷，高效，超前，可靠的诊断方法，实验以ＤＭＢＡ（二甲基苯并蒽）诱导的金地鼠颊囊致癌模型为对象，以ＰＣＮＡ（增殖细胞核抗原）ＢｒｄＶ（溴脱氧尿嘧啶）等免疫组化技术为手段；以光镜观察诊断为对照，对三种观察结果进行相关分析，发现三者间有高度显著的相关性，证实了免疫组化方法在癌变诊断中有一定的参考价值，实验还发现ＰＣＮＡ和ＢｒｄＵ检测结果比较每天敏感，简捷，     

增殖细胞核抗原在口腔鳞状细胞癌中的表达

采用鼠抗ＰＣＮＡ单克隆抗体，ＡＢＣ法，对１１例正常口腔粘膜及２９例口腔鳞癌进行了染色。结果表明：正常粘膜仅在基底层中有少量的ＰＣＮＡ阳性细胞，其阳性分级为１级。而鳞癌阳性分级主要是３～４级，随着组织学分级的升高，ＰＣＮＡ阳性表达率也相应增高。经统计学分析，正常组织与鳞癌之间及鳞癌各级均有非常显著差异（Ｐ＜００１）。提示：ＰＣＮＡ的表达与口腔鳞癌组织学分级有关。     

Proliferative activity of calcifying odontogenic cysts as evaluated by proliferating cell nuclear antigen labeling index

The calcifying odontogenic cyst (COC) presents with diverse hlstologlcal features; thus, several subclasslfl-cations have been proposed. To evaluate the slgnlficance of the various histological features and subtypes of COC from the perspectlve of proliferative activity, the proliferating cell nuclear antigen (PCNA) labellng index (LI; the percentage of positive nuclei) was assessed immunohistochemlcally in 25 cases of COC (21 benign and four malignant). All of the benign cases were of the cystic variety and further subclas-sified into non-proliferative subtype (NPS; four cases); proliferative subtype (PS; eight cases); and COC associated with odontoma (COCaO, nlne cases). The PCNA U of the mallgnant COC (65.2 ± 5.6) was slgnlflcantly higher than that of the benlgn COC (11.6 ± 9.0; P = 0.002). Non-proliferative subtype (6.8 ± 2.8) showed the lowest PCNA LI and PS (17.2 ± 11.2) the highest of among the three subtypes of benign cystic COC (P = 0.028). In nine cases of COCaO, six showed epithelial lining of the non-proliferative type as NPS and the other three had lining wlth proliferative features as PS. The PCNA LI of the latter COCaO group (14.3 ± 6.6) was significantly higher than that of the former (6.1 ± 4.3; P = 0.05), as Seen between PS and NPS. These results demonstrate that PCNA LI is a possible parameter for differentiating mallgnant COC from benign COC and, whatever the subtypes, the proliferative features In the lining are the main factor influencing the prollferatlng actlvity of COC.     

甲型肝炎早期实验诊断的研究

以人胚肺二倍体细胞分离出的甲型肝炎病毒作抗原,以马抗人IgM经纯化后的抗体为捕捉抗体,以纯化甲型肝炎恢复血清为特异性抗体,过碘酸钠法标记酶,以OPD为底物制成抗-HAV IgM诊断试剂盒,建立了ELISA捕捉法检测抗-HAV IgM,结果:甲型肝炎暴发点84例均为阳性,流行性出血热及健康成人各30例均为阴性。322例急性黄疸型肝炎住院患者中192例阳性(占59.6%)。12例第一病日患者11例阳性(占91.6%),证明该方法对甲型肝炎早期诊断有重要价值。     

The significance of HIV antigen and HIV antibodies specific against envelope and core proteins in serum of HIV-infected hemophiliacs

Serial serum samples from the 21 HIV-infected Norwegian hemophiliacs have been assayed for the presence of HIV antigen and antibodies to HIV specific for the core protein p24 and the envelope protein gp41. HIV antigen was detected in 4 patients, of whom 3 have developed AIDS to date. HIV antigen appeared in serum 10 to 24 months before the diagnosis in these patients. Antibodies to gp41 was a constant finding. Antibodies to p24 disappeared from the serum in 1 patient who developed AIDS and from 3 persons free of symptoms related to the HIV infection. The detection of HIV antigen in HIV antibody-positive hemophiliacs appears to be of considerable prognostic significance.