McAbGB_2识别的乳腺癌血清抗原水平观察

为探明乳腺癌患者由ＭｃＡｂＧＢ２识别的乳腺癌血清抗原（以下简称血清ＭｃＡｂＧＢ２抗原）水平，应用ＥＬＩＳＡ方法，用抗人乳腺癌血清抗原ＭｃＡｂＧＢ２观察了５０例正常人和６０例乳腺癌患者血清ＭｃＡｂＧＢ２抗原水平。结果：１７例乳腺癌患者术前的血清ＭｃＡｂＧＢ２抗原水平与正常人之间有显著性差异（Ｐ＜０．００１），其阳性符合率达８８．２％（１５／１７）。其中８例乳腺癌患者的血清ＭｃＡｂＧＢ２抗原水平由术前的５７．５±５１．３ｕ／ｍｌ降至术后的２０．６±４．９８ｕ／ｍｌ。４３例在化疗中的乳腺癌术后患者的血清ＭｃＡｂＧＢ２抗原多处于正常水平。由此提示血清ＭｃＡｂＧＢ２抗原可作为乳腺癌标志物，用于乳腺癌的血清学诊断、治疗及预后监测等     

抗人乳腺癌血清抗原单克隆抗体GB2的制备及鉴定

采用人乳腺癌血清抗原免疫的ＢＡＬＢ／ｃ小鼠脾细胞与小鼠骨髓瘤细胞ＳＰ２／０融合，经ＥＬＩＳＡ筛选，有限稀释及克隆后获得了一株稳定分泌特异抗体的杂交瘤细胞株，命名为单克隆抗体ＧＢ２（ＭｃＡｂＧＢ２）。ＭｃＡｂＧＢ２经硫酸铵及ＤＥＡＥ纤维素纯化后效价达１∶２×１０６，经琼脂糖免疫双扩证明ＭｃＡｂＧＢ２属小鼠ＩｇＧ１亚类。免疫印迹结果表明ＭｃＡｂＧＢ２识别的抗原呈两条区带，分子量分别为１１６ｋｄ及４５ｋｄ，分布于乳腺癌组织及血清中。     

抗人乳腺癌血清抗原单克隆抗体GB2的制备及鉴定

采用人乳腺癌血清抗原免疫的ＢＡＬＢ／ｃ小鼠脾细胞与小鼠骨髓瘤细胞ＳＰ２／０融合，经ＥＬＩＳＡ筛选，有限稀释及克隆后获得了一析玢泌特异抗体的杂交瘤细胞株，命名为单克隆抗体ＧＰ２（ＭｃＡｂＧＢ２）。ＭｃＡｂＧＢ２经硫酸铵及ＤＥＡＥ纤维素纯化后效价达１：２×１０＾６，经琼脂糖免疫双扩证明ＭｃＡｂＧＢ２属小鼠ＩｇＧ１亚类。免疫印迹结果表明ＭｃＡｂＧＢ２识别的抗原呈两条区带，分子量分别为１１６ｋｄ及４５ｋｄ     

分泌抗人喉癌单克隆抗体杂交瘤细胞株的建立及临床意义

用喉癌抗原免疫ＢＡＬＢ／Ｃ小鼠的脾细胞与ＮＳ－１小鼠骨髓瘤细胞融合，经筛选获得４株分泌抗人喉癌单克隆抗体（ＭｃＡｂ）的杂交瘤细胞株，均收集到腹水型ＭｃＡｂ，并验证其滴度高，纯化效果好，特异性强。用其中３株ＭｃＡｂ混合，对１２４例喉癌及４０例正常人血清予以检测，结果喉癌患者血清中喉癌相关抗原水平明显高于对照组，Ｐ＜０．０１，且随病情的加重，喉癌相关抗原水平呈上升趋势。故本室建立的抗人喉癌混合ＭｃＡｂ可作为一种较为理想的肿瘤标记物，为喉癌的诊断、病情监测、预后判定提供了一种敏感、特异的测试手段。     

人甲状腺过氧化物酶的纯化及与微粒体抗体的关系

用胆酸钠增溶、硫酸铵盐析、差速离心及免疫亲和层析法从甲状腺细胞膜成分中提纯ｈＴＰＯ，纯化ｈＴＰＯ的比生物活性提高了４３８．７倍，其ＲＺ值为０．２５．ＳＤＳ－ＰＡＧＥ于１０５ＫＤ和１０７ＫＤ处呈现两条条带。以固相Ｉｏｄｏｇｅｎ法标记ｈＴＰＯ，建立放免法测定２９例ＡＩＴＤ患者ＭｃＡｂ阳性血清的ＴＰＯＡｂ结合率为１６．３％±６．３％，而１１例正常人ＭｃＡｂ阳性血清的结合率为３．４％１４．７％，ＭｃＡｂ与ＴＰＯＡｂ结合率的测定值呈显著相关（ｒ＝０．７３，Ｐ＜０．００１）；稀释试验提示ＭｃＡｂ可与ｈＴＰＤ结合且存在剂量依赖性。因此ＡＩＴＤ患者的ＭｃＡｂ所针对的微粒体抗原即为ｈＴＰＯ，而ＭｃＡｂ即为ＴＰＯＡｂ。     

血吸虫循环免疫复合物解离后抗原及抗体的免疫反应性研究

将血吸虫循环免疫复合物（ＣＩＣ）用新方法解离后，特异性测定其上的抗原（Ａｇ）及抗体（Ａｂ），并以可溶性血吸虫卵抗原（ＳＥＡ）、多克隆、单克隆抗体（Ｐｃａｂ、ＭｂＡｂ）用间接法对比测定感染者血清中循环抗原（ＣＡｇ）及循环抗体（ＣＡｂ）通过１０２例日本血吸虫病人及４５名健康人ＣＩＣ解离后和血清的特异性对比测定结果表明，ＣＩＣ解离后的Ａｇ及Ａｂ水平（ＯＤ均值）测定，显示满意的特异性和敏感性，与用抗ＳＥＡ－和抗ＣＣＡ－ＭｃＡｂ测定ＣＡｇ水平（ＯＤ均值）相比较，具有优于或相似的检测效果，采用抗ＳＥＡ－ＰｃＡｂ和ＳＥＡ测定血谱中的ＣＡｇ及ＣＡｂ水平（ＯＤ均值）时，均取得很好效果，且其ＣＡｇ水平均显著地高于用２种ＭｃＡｂ检测ＣＡｇ的水平。上述制剂测定ＣＩＣ解离后及血清中的Ａｇ、Ａｂ及ＣＡｇ、ＣＡｂ，可起着互补诊断的效果。本项研究结果可为今后选用适宜的血吸虫病血清学诊断的检测途径，提供重要的参考依据。     

凋亡相关基因bcl-2基因族在乳腺癌中的预后作用

探讨癌基因ｂｃｌ－２及其基因族成员ｂａｘ、ｂｃｌｘ的表达水平与乳腺癌生物学行为的关系及对乳腺癌预后的意义。方法运用免疫组化方法对９１例乳腺癌石蜡组织切片ｂｃｌ－２、ｂａｘ蛋白表达水平进行检测,运用Ｎｏｒｔｈｅｒｎ印迹法检测１６例新鲜乳腺癌组织标本ｂｃｌｘｌｍＲＮＡ表达水平。结果９１例乳腺癌病例中,ｂｃｌ２阳性率６５．９％（６０／９１）,ｂａｘ阳性率６４．８％（５９／９１）。ｂｃｌ２、ｂａｘ蛋白表达水平与凋亡指数（ＡＩ）、病理组织学分级、腋淋巴结转移情况、术后局部复发与转移相关,ｂｃｌ２表达与雌激素受体（ＥＲ）水平正相关。ＡＩ与ｂｃｌ２、ｂａｘ蛋白表达水平相关。对影响乳腺癌术后无病生存率的单因素分析结果显示,ｂｃｌ２、ｂａｘ蛋白表达水平具有预后作用;Ｃｏｘ’ｓ模型多因素分析结果,ｂｃｌ２蛋白表达水平是独立的预后指标。１６例乳腺癌新鲜标本中,５０％（８／１６）ｂｃｌｘｌｍＲＮＡ高表达,其表达水平与ｂｃｌ２蛋白表达、腋淋巴结转移相关。结论凋亡相关基因ｂｃｌ２、ｂａｘ、ｂｃｌｘｌ表达失调可导致乳腺癌细胞凋亡调控的紊乱,使腋淋巴结转移程度增加,ｂｃｌ２蛋白表达水平在乳腺癌中有独立的预后价值     

凋亡相关基因bcl—2基因族大乳腺癌中的预后作用

目的 探讨癌基因ｂｃｌ－２及其基因族成员ｂａｘ、ｂｃｌ－ｘ的表达水平与乳腺癌生物学行为的关系及对乳腺癌预后的意义。方法 运用免疫组化方法对９１例乳腺癌石蜡组织切片ｂｃｌ－２、ｂａｘ蛋白表达水平进行检测,运用Ｎｏｒｔｈｅｒｎ印迹法检测１６例新鲜乳腺癌组织标本ｂｃｌ－ｘｌｍＲＮＡ表达水平。结果 ９１例乳腺癌病例中,ｂｃｌ－２阳性率６５．９％,ｂａｘ阳性率６４．８％。ｂｃｌ－２、ｂａｘ蛋白表达水平与凋亡     

风疹病毒单克隆抗体的制备

目的 制备用于研究风疹病毒（ＲＶ）生物学特性及血清学应用的特异性ＭｃＡｂ。方法 ＨＶ抗原免疫ＢＡＬＢ／ｃ小鼠,无菌取其脾细胞与小鼠骨髓瘤细胞ＳＰ２／０融合建立杂交瘤细胞,并对杂交瘤细胞产生抗体特性及相应ＭｃＡｂ进行了分析。结果 成功建立了三株稳定分泌ＭｃＡｂ的杂交瘤细胞５Ｄ５、５Ｇ５、２Ｆ７,染色体数目均大于９８条;相应ＭｃＡｂｓ均属ＩｇＧ１。与纯化ＲＶ抗原有强阳性反应。结论 该法可产出高特异性、高亲和力ＲＶ特异性ＭｃＡｂ。     

筑巢式PCR检测慢性粒细胞白血病bcr—abl mRNA

目的：观察慢性粒细胞性白血病（ＣＭＬ）患者ｂｃｒ／ａｂｌ ｍＲＮＡ的表达情况。方法：应用逆转录筑巢式聚合酶链反应（Ｎｅｓｔｅｄ ＰＣＲ）检测了３２例ＣＭＬ患者的ｂｃｒ／ａｂｌ ｍＲＮＡ。结果：３０例检测到ｂｃｒ／ａｂｌ ｍＲＮＡ,其中１９例检测到ｂｃｒ ｅｘｏｎ３／ａｂｌ ｅｘｏｎ２ ｍＲＮＡ,７例表达ｂｃｒ ｅｘｏｎ２／ａｂｌ ｅｘｏｎ２ ｍＲＮＡ。４例同时表达这两种ｍＲＮＡ。结论：Ｎｅｓｔｅｄ     

肺癌患者血清内皮抑素水平对肺癌早期诊断价值的研究

目的探讨肺癌患者不同临床分期、组织学类型血清内皮抑素的水平以及化疗前后其水平的变化,从而判断血清内皮抑素水平对肺癌的早期诊断和预后的价值。方法对40例肺癌患者、20例良性肺病患者和20例正常人用ELISA方法检测血清内皮抑素。对不同临床分期、病理类型及化疗前后血清内皮抑素的水平进行分析。结果(1)肺癌组血清内皮抑素水平(10．71±9．99)ng／ml高于良性肺病组(4．79±1．23)ng／ml和正常对照组(4．51±1．14)ng／ml,差异有统计学意义(P<0．01);(2)Ⅰ、Ⅱ期肺癌患者血清内皮抑素的水平高于Ⅲ期,差异有统计学意义(P<0．05)。(3)肺癌患者不同病理类型血清内皮抑素水平的比较差异无统计学意义。(4)肺癌患者化疗后血清内皮抑素水平明显高于化疗前,差异有统计学意义(P<0．05)。结论(1)血清内皮抑素可能成为肺癌早期诊断的参考指标;(2)血清内皮抑素水平的动态变化可作为化疗疗效观察和预后的评价指标。     

Immune function of cancer patients with spleen-deficiency syndrome

According to this study, the immunological function was aberrant in cancer patients with Spleen-deficiency syndrome. The TII cell in normal persons (n = 26) was 30, 86 +/- 9.70% (means +/- S) and in these cases (n = 43) was 22.62 +/- 9.92%, P less than 0.002. The cytotoxicity of NK cell in patients (n = 59) was 17.65 +/- 10.58%, in normal controls (n = 43) was 25.51 +/- 14.10%. The combining ability of NK cell in patients (n = 48) was 39.11 +/- 19.43%, the normal persons (n = 41) was 55.88 +/- 17.94%. It showed that the immune function of the cancer patients with Spleen-deficiency syndrome were markedly lower than that of normal persons. The serum IgA in saliva of patients (n = 37) was 0.44 +/- 0.17 microgram/ml. It was much higher than that of normals' (n = 24, 0.30 +/- 0.06 microgram/ml), P less than 0.001. Some patients' NK cell function and the level of level of SIgA in saliva were recovered to normal after treatment of Shengxue Tang which could strengthen the Spleen and replenish the Kidney. These studies proved that the TCM played an important role for modulating immune function in treating cancer patients.     

血清血管内皮生长因子C水平及癌组织中环氧化酶2的表达与乳腺癌淋巴道转移的关系

目的 探讨乳腺癌患者术前血清血管内皮生长因子C(sVEGF-C)水平、组织中环氧化酶2(COX-2)表达与乳腺癌淋巴道转移的关系.方法 采用ELISA法检测68例乳腺癌患者、35例乳腺良性病术前以及12名健康女性sVEGF-C水平.免疫组化SP法检测相应乳腺癌患者组织标本中COX-2表达及淋巴管密度(LMVD);分析sVEGF-C、COX-2及LMVD表达之间的关系.结果 (1)乳腺癌患者术前sVEGF-C水平(49±8)pg/ml显著高于健康女性志愿者(5±3)pg/ml和乳腺良性疾病者(8±4)pg/ml;(2)乳腺癌组织COX-2阳性表达率(44.1%)显著高于乳腺良性疾病(11.4%)(P=0.002);LMVD主要位于癌旁组织,癌巢中少有表达;癌旁组织中LMVD(8.7±4.7)较乳腺良性疾病组织中表达(1.8±1.7)显著增多(P=0.002);(3)在乳腺癌患者sVEGF-C水平增高者中,不但COX-2的表达显著高于sVEGF-C水平正常者COX-2的表达(P=0.024),而且LMVD(8.7±3.9)也显著高于sVEGF-C正常者的LMVD(5.6±3.3)(P=0.039);(4)乳腺癌患者COX-2表达阳性组LMVD(9.5±3.7)明显高于COX-2表达阴性组LMVD(6.0±3.8)(P=0.012);(5)在腋淋巴结转移阳性患者中,sVEGF-C水平、COX-2及LMVD表达均高于腋淋巴结转移阴性组(P值分别为0.025,0.022和0.002).结论 sVEGF-C、COX-2及LMVD在乳腺癌的淋巴道转移中有重要作用,可作为估计乳腺癌有无淋巴结转移的参考指标.     

脑膜瘤患者手术前后血清表皮生长因子受体的测定

目的探讨脑膜瘤患者手术前后外周血表皮生长因子受体(EGFR)表达含量的临床意义。方法采用ELISA分析53例脑膜瘤患者(42例肿瘤完全切除和11例肿瘤次全切除)手术前后的血清EGFR含量。28例正常献血者为对照组。结果53例脑膜瘤患者手术前血清EGFR含量(352.93±66.18)fmol/ml明显高于对照组(159.11±40.50)fmol/ml(P<0.0001);手术后(220.74±70.63)fmol/ml明显低于术前组(P<0.001)。其中42例肿瘤完全切除患者中38例的EGFR(191.20±32.13)fmol/ml和4例伴有瘤周水肿患者的EGFR(248.75±10.31)fmol/ml明显低于术前组(P<0.001);11例肿瘤次全切除患者的EGFR(322.14±89.53)fmol/ml与术前组比较无显著性差异(P>0.5)。结论92.16%脑膜瘤患者随着肿瘤切除程度的增高而血清EGFR含量越发降低。提示人脑膜瘤细胞可能自主产生EGFR,血清EGFR测定对脑膜瘤患者手术后追踪可能具有临床意义。     

Establishment and preliminary application of spermidine radioimmunoassay with 125I-labelled monoclonal antibody and solid phase antigen.

Purified anti-spermidine monoclonal antibody was labelled with radioactive iodine by the Iodogen method and spermidine-bovine serum albumin (SPD-BSA) conjugate was used to coat polystyrene beads as solid phase antigen. The new solid phase 125I-labelled spermidine radioimmunoassay (RIA) depends on the competition between spermidine in the sample and the solid phase antigen for the limited amount of 125I-labelled monoclonal antibody. The sensitivity of this assay was 10 ng/ml higher than that of liquid RIA for spermidine with 14C-labelled spermidine. The coefficients of variation (CV) within and among batches were 4% and 13% respectively. The sample-batch capacity was increased from 20 (liquid RIA with 14C-labelled spermidine) to 150-200 by using this method. Because of its simplicity, the solid phase RIA kit is very convenient for population survey. This RIA could be used to determine spermidine in saliva for the diagnosis of precancerous lesions. In a preliminary study saliva spermidine levels in different populations were measured among 130 normal subjects, 202 esophageal epithelial hyperplasia cases treated with anti-tumor B for 5 years, 207 esophageal epithelial hyperplasia cases as control, and 55 esophageal cancer patients. The levels were 1,795 +/- 1,481, 3,470 +/- 6,981, 9,753 +/- 17,641 and 18,090 +/- 21,509 ng/ml, respectively, with the saliva spermidine levels in precancerous and cancer patients being significantly higher than that of normal subjects (P less than 0.001); the level in patients treated with anti-tumor B was significantly lower than that of controls (P less than 0.001). This decreased saliva spermidine content was coincident with the 47.3% reduction of canceration rate seen in precancerous patients after a 5-year treatment with anti-tumor B.     

Vascular endothelial growth factor expression in serum of patients with hepatocellular carcinoma

Objectives To determine the pre-therapeutic serum level of vascular endothelial growth factor (VEGF) in patients with hepatocellular carcinoma (HCC) and to elucidate the relation between the serum level and clinical characteristics and metastasis of HCC. Methods One-hundred and fifteen HCC patients, 40 patients with benign liver lesions, and 30 healthy control subjects were included in this study. The serum VEGF level was measured with the quantitative sandwich enzyme linked immunosorbent assay (ELISA, R&amp;D systems). Results The serum VEGF levels in the HCC group (465.62±336.24 pg/ml) was significantly elevated as compared with those in patients with benign liver lesions (159.54±120.58 pg/ml) and those in normal controls (123.53±51.84 pg/ml). The VEGF levels were not significantly different between the patients with benign liver lesions and the normal controls. The serum VEGF concentration showed a positive rate of 77.4%, 25%, and 3.3% in the HCC patients, benign liver lesion patients and normal controls, respectively. In the 115 HCC patients, the serum VEGF levels in patients with portal vein (PV) emboli (n=26, 582.76±441.89 pg/ml), with metastasis (n=43, 548.29±438.57 pg/ml) or with large HCC lesions (≥5 cm in diameter) (n=69, 554.43±369.99 pg/ml) were significantly higher than those without PV-emboli (n=89, 431.39±292.84 pg/ml), without metastasis (n=72, 416.24±247.27 pg/ml) or with small HCC lesions (n=42, 328.67±227.47 pg/ml). The serum VEGF levels in stage Ⅰ, Ⅱ, Ⅲ, Ⅳa and Ⅳb HCC patients were 340.6 pg/ml, 451.55±307.84 pg/ml, 397.44±257.18 pg/ml, 486.10±397.73 pg/ml and 647.93±344.56 pg/ml, respectively. Conclusion The pre-therapeutic serum VEGF levels in HCC patients appear to reflect the disease’s potential activity of vascular invasion and metastasis.     

乳腺癌患者血清CA15－3的临床意义

用ＩＲＭＡ法检测４２例乳腺癌患者术前和术后第２周血清ＣＡ１５－３值，并结合临床病理资料进行分析。结果，血清ＣＡ１５－３水平与淋巴结转移状态显著相关（Ｐ＜０．０１）；术前术后ＣＡ１５－３水平差异有显著性（Ｐ＜０．０５）；其中３例有远处转移的病人，其ＣＡ１５－３值均高于１００ｕ／ｍｌ。因而认为，血清ＣＡ１５－３水平可作为乳腺癌转移的辅助性诊断手段，并用于估计肿瘤转移的潜在可能性，为监测肿瘤复发提供有力的佐证     

胰液糖抗原19—9含量对胰腺癌的诊断意义

糖抗原19-9(CA 19-9)是从结肠癌细胞株SW 1116分离的抗原成分,其血清含量在许多消化道肿瘤均增高,因而被认为是一种胃肠道肿瘤相关抗原。本文探讨CA19-9在胰腺癌诊断中的意义。对象与方法CA19-9测定 用单克隆抗体固相免疫放射药盒,由美国CENTOCOR公司惠赠。研究对象 432名,其中正常人71名(本院工作人员、学生及献血员),良性消化系疾病116例(包括慢性胃炎、溃疡病、总胆管结石及慢性胰腺炎),胰腺癌44例,壶腹癌22例,胆管癌19例,胃癌98例,     

血清T-PSA和F-PSA检测在乳腺癌诊断中的意义

目的 :检测术前乳腺癌患者血清中T PSA和F PSA的含量 ,探讨其和乳腺癌临床生物学行为的关系及在诊断中的价值。方法 :利用微粒子酶免分析法 ,检测 85名女性乳腺癌患者和 30名健康女性血清中T PSA和F PSA的含量。结果 :指标的最低检测值为 0 .0 1ng/ml。乳腺癌患者血清中F PSA水平明显高于健康女性 (P <0 .0 5 ) ,F PSA优势患者占 3.76 % ,而健康女性中仅占 3.3%。T PSA和F PSA的阳性判断值 (cut offvalue)分别为 0 .36ng/ml和0 .0 2ng/ml,女性乳腺癌患者的T PSA和F PSA阳性率分别为 2 3.5 %和 2 7.1%。T PSA阳性患者中 ,淋巴结转移者较多 (70 % ,P >0 .0 5 ) ,而F PSA优势患者中 ,早期 (Ⅰ、Ⅱ期 )发病率较高 (6 8.8% ,P >0 .0 5 )。结论 :虽然F PSA占优势鉴别乳腺癌的敏感性较低 ,但特异性较高。早期乳腺癌有F PSA占优势的趋势。血清检测T PSA和F PSA对乳腺癌的诊断及预后判断有一定意义     

P-钙粘蛋白在乳腺癌患者血清中的表达及其临床意义

目的检测P-钙粘蛋白在乳腺癌患者血清中的表达及其临床意义。方法收集乳腺癌患者60例、乳腺良性肿瘤患者46例及健康对照组30名,采用酶联免疫吸附试验(ELISA)法检测各组研究对象血清中P-钙粘蛋白的表达水平,并分析血清中P-钙粘蛋白水平与乳腺癌病理指征的关系。结果乳腺癌组血清P-钙粘蛋白水平为(36.98±4.53)ng/ml,明显高于乳腺良性肿瘤组(20.24±6.32)ng/ml和对照组(10.31±1.85)ng/ml,差异有统计学意义(P0.05)。乳腺癌临床分期Ⅲ期患者血清P-钙粘蛋白水平为(48.49±3.87)ng/ml,明显高于临床Ⅰ~Ⅱ期患者(26.35±2.04)ng/ml,差异有统计学意义(P0.05)。分化程度Ⅲ级患者血清P-钙粘蛋白水平为(41.13±2.35)ng/ml,明显高于分化程度Ⅰ~Ⅱ级患者(26.35±2.04)ng/ml,差异有统计学意义(P0.05),淋巴结转移患者血清P-钙粘蛋白水平为(42.26±3.32)ng/ml,明显高于无淋巴结转移组(28.73±1.48)ng/ml,差异有统计学意义(P0.05)。结论检测血清P-钙粘蛋白水平对乳腺癌的诊断、病情监测等具有一定的参考作用。