NSCLC术后病人外周血MUC1 mRNA和CK19 mRNA表达及其意义

目的探讨非小细胞肺癌（NSCLC）术后外周血微转移的基因诊断方法，并分析黏蛋白1（MUC1）mRNA、角蛋白19（CK19）mRNA作为肺癌微转移检测分子标志物的可行性。方法应用逆转录聚合酶链反应（RT—PCR），对60例NSCLC病人（肺癌组）的外周血MUC1 mRNA、CK19 mRNA表达情况进行检测；以10例肺良性病变病人及15例健康人的外周血标本作对照。结果肺癌组外周血MUC1 mRNA阳性表达21例（35%），CK19 mRNA阳性表达18例（30％）；对照组MUC1 mRNA和CK19 mRNA表达均为阴性，两组比较差异均有显著性（x^2=7．292、5．921，P〈0．01、0．05）。结论MUC1、CK19基因均可作为RT—PCR法检测NSCLC病人淋巴结微转移的分子标志物，两者联合检测可能有助于肺癌转移早期诊断。     

局部寡核苷酸诱导小鼠毛囊角蛋白17基因改变的基因治疗研究

目的 研究体内角质形成细胞基因修改的可能性。方法 通过RNA-DNA寡核苷酸(RDO)或单链DNA寡核苷酸(ssODN)局部皮肤注射的方法，诱导毛囊角蛋白17(K17)(R94P)显性突变。注射的K17A-RD0或K17A-ssODN含有一个单碱基错配(CGC到CCC)，从而引起该氨基酸的替换。注射的混合物由上述寡核苷酸和阳离子脂质体组成。分别于小鼠出生后第2天和第5天进行皮内注射。第8天取皮肤活检。结果 组织学检查发现注射K17A-RD0的7只小鼠中有5只和注射K17A-ssODN的5只小鼠全部有毛发生长明显异常。其主要表现为毛囊内毛干在皮脂腺水平扭曲盘绕或断裂，个别毛囊毛球部碎裂，黑素颗粒滴落到周围真皮中，有些切片中还可见表皮囊肿。注射部位生长期Ⅳ～Ⅵ期毛囊数减少50％。阴性对照组均未显示明显毛发生长异常。从组织切片上分离异常毛囊并提取DNA,PCR扩增后DNA测序和非限制性内切酶酶切证实，K17基因由正常的CGC变成了CCC。结论 局部注射K17A-RD0或K17A-ssODN可以导致特异性靶基因K17的定点突变，引起可见的毛发生长异常。     

人源性抗角蛋白Fab抗体脂质体的制备

目的制备人源性抗角蛋白Fab抗体(HAKFA)脂质体,检测脂质体的体外释放及包裹和游离抗体的活性,探索HAKFA可行的应用途径。方法采用逆相蒸发与钙融合联合法制备HAKFA脂质体,在透射电镜下观察脂质体颗粒的性状及大小,应用紫外分光光度计检测抗体的包封率,ELISA检测包裹前后的抗体活性。结果本方法制备的脂质体呈现圆形、粒径大小较一致,约50~100nm;HAKFA脂质体包封率达到(60.3±12.7)%;HAKFA脂质体呈现缓释特性并且抗体依然保持良好的抗原结合活性。结论逆相蒸发与钙融合法为制备HAKFA脂质体较理想的方法。     

Cyclic AMP-dependent protein kinase in calf-snout epidermis

Summary The biochemical characteristics of cyclic AMP-dependent protein kinase in calf-snout epidermis were investigated. The activity of cyclic AMP-dependent protein kinase was higher in the lower layer than the upper layer of epidermis. The supernatant of homogenates of the lower layer of calf-snout epidermis was fractionated by DEAE-cellulose chromatography and contained two major peaks of protein kinase activity stimulated by cyclic AMP. This chromtographic pattern is similar to that referred to as Type I and Type II of cyclic AMP-dependent protein kinase in bovine muscle. Both peaks of cyclic AMP-dependent protein kinase in calf-snout epidermis could phosphorylate keratin polypeptides in vitro. The phosphorylation reaction was activated by cyclic AMP and inhibited by a heat-stable inhibitor of cyclic AMP-dependent protein kinase. When Type II enzyme of cyclic AMP-dependent protein kinase was incubated with [-³²P]ATP in the absence of substrates, such as histone or keratin polypeptides, the 54,000 dalton protein was phosphorylated and this autophosphorylation was inhibited by the addition of 10 M cyclic AMP. These results suggest that cyclic AMP-dependent protein kinase in calf-snout epidermis has properties similar to those in bovine muscle and plays an important role in the phosphorylation of keratin polypeptides in calf-snout epidermis.     

Twist基因蛋白表达在甲状腺乳头状癌诊断与鉴别诊断中的意义

目的探讨Twist基因蛋白在甲状腺乳头状癌中的表达状况,寻找更特异的标记物,用于甲状腺乳头状癌（PTC）与滤泡状腺瘤（FA）和良性乳头病变（BPL）的鉴别诊断。方法以50例PTC为研究组,以48例FA和47例BPL作对照组。在自制组织芯片上行免疫组织化学（SP法）标记,检测Twist基因蛋白和HBME-1,并以CK19进行对照。结果3种指标（Twist、HBME-1、CK19）阳性表达率：在PTC组分别为100％（48／48）、94．0％（47／50）和78．0％（39／50）;在FA组分别为0、6．7％（3／45）和0;在BPL组分别为7．0％（3／43）、2．1％（1／47）和0。PTC组分别与FA和BPL组比较差异均有统计学意义（P=0．000）。3种指标在鉴别诊断甲状腺良、恶性病变的灵敏度分别为：100％、96．4％和97．7％,特异度分别为：94．0％、95．7％和95．1％,准确度分别为：78．0％、100％和91．9％。结论Twist可应用于辅助诊断PTC,并可应用在对FA或BPL的鉴别诊断。     

Phenotypic plasticity of neoplastic ovarian epithelium: unique cadherin profiles in tumor progression

The mesodermally derived normal ovarian surface epithelium (OSE) displays both epithelial and mesenchymal characteristics and exhibits remarkable phenotypic plasticity during post-ovulatory repair. The majority of epithelial ovarian carcinomas (EOC) are derived from the OSE and represent the most lethal of all gynecological malignancies, as most patients (approximately 70%) present at diagnosis with disseminated intra-abdominal metastasis. The predominant pattern of EOC metastasis involves pelvic dissemination rather than lymphatic or hematologic spread, distinguishing EOC from other solid tumors. Acquisition of the metastatic phenotype involves a complex series of interrelated cellular events leading to dissociation (shedding) and dispersal of malignant cells. A key event in this process is disruption of cell-cell contacts via modulation of intercellular junctional components. In contrast to most carcinomas that downregulate E-cadherin expression during tumor progression, a unique feature of primary well-differentiated ovarian cancers is a gain of epithelial features, characterized by an increase in expression of E-cadherin. Subsequent reacquisition of mesenchymal features is observed in more advanced tumors with concomitant loss of E-cadherin expression and/or function during progression to metastasis. The functional consequences of this remarkable phenotypic plasticity are not fully understood, but may play a role in modulation of cell survival in suspension (ascites), chemoresistance, and intraperitoneal anchoring of metastatic lesions.