抗γ 精浆蛋白特异性单链抗体的RTS表达及其生物学活性的检测

目的〖HT5"SS〗： 制备人精浆蛋白特异性单链抗体,并检测其生物活性与靶向性。〖HT5W〗方法：〖HT5"SS〗 应用快速翻译系统（RTS）表达γ精浆蛋白特异性单链抗体,制备E4B7ScFv肽段和纳米颗粒偶联物,应用免疫沉淀反应、Western Blotting法、免疫荧光染色分析和细胞实验,研究该抗体进入前列腺癌细胞的活性。〖HT5W〗结果：〖HT5"SS〗成功地制备了抗γ精浆蛋白特异性单链抗体纳米颗粒偶联物。该抗体可与前列腺癌细胞特异性结合,并可在15 min内将纳米磁珠带入前列腺癌细胞内,进入癌细胞的磁球数量随着细胞培养时间的延长而增多,对照实验显示该偶联物不能进入其它组织肿瘤细胞。〖HT5W〗结论：〖HT5"SS〗制备的抗γ精浆蛋白特异性单链抗体生物活性与靶向性均较好,在前列腺癌的早期核磁共振显像诊断与靶向治疗上具有潜在的应用价值。     

乳腺癌单链抗体制备及其在荷人乳腺癌裸鼠中的放免显像

目的：制备抗乳腺癌单链抗体,用于荷瘤裸鼠的放免显像研究。方法：提取乳腺癌膜抗原,制备抗乳腺癌单克隆抗体M4G3,用Pharmacia噬菌体单链抗体制备系统得到可溶性具有抗乳腺癌活性的ScFv抗体,利用环DTPA法标记核素99^mTc,进行体外培养的乳腺癌细胞系MCF-7和荷瘤小鼠体内定位显像。结果：免疫组化显示单链抗体与乳腺癌膜抗原有较强的亲和力,并在体外培养的乳腺癌细胞及荷瘤小鼠体内显像成功,且在抗体注射第3-5天显像最清晰。结论：提示M4G3的ScFv可在乳腺癌的放射免疫显像和定位诊断研究中发挥作用,值得进一步研究。     

钆-叶酸-四氧化钴铁磁性纳米粒子对卵巢癌细胞靶向性能的影响

[目的]探讨钆-叶酸-四氧化钴铁磁性纳米粒子通过靶向结合卵巢癌COC1细胞,在核磁共振成像时对卵巢癌组织对比度的性能影响.[方法]利用透射电镜观察纳米粒子靶向进入卵巢癌细胞的过程,采用荧光光谱法测定不同浓度下纳米粒子进入细胞的定量,MTT法检测纳米粒子的细胞毒性,再通过核磁共振成像技术分析不同浓度纳米粒子对T1层面成像、T2层面成像的作用.[结果]纳米粒子能够靶向进入卵巢癌COC1细胞,并呈剂量依赖性;在进入细胞48h内未显示出细胞毒性,并能够在T1、T2两个层面增加核磁共振成像的组织对比度.[结论]钆-叶酸-四氧化钴铁磁性纳米粒子能够增加卵巢癌核磁共振成像组织的对比度,提高其分辨率,有可能成为一种既能改变T1成像,又能改变T2成像分辨率的新的靶向磁性造影剂.     

人黑素瘤单链抗体-量子点纳米探针的制备及其与黑素瘤细胞特异性结合

目的：制备抗人黑素瘤细胞单链抗体与碲化镉量子点(CdTe guantun dot, CdTe)连接的纳米探针,观察其对恶性黑素瘤细胞的特异性结合效果。方法：将抗人黑素瘤神经节苷脂单链抗体（antihuman melanoma ganglioside single chain variable fragment antibody,GD/ScFvMEL）基因克隆到pET32a（+）载体中,然后在BL21（DE3）细菌中进行诱导表达,采用变性法纯化表达的蛋白,再用改良的透析法复性目的蛋白,SDSPAGE分析表达的GD/ScFvMEL抗体蛋白。制备的GD/ScFvMEL抗体与量子点连接制备成纳米探针GD/ScFvMELQDs,然后与黑素瘤A375细胞株及胃癌MGC803细胞株孵育,观察纳米探针与肿瘤细胞结合的特异性。结果：PCR、双酶切和序列测定证实重组子的拼接完全正确,SDSPADE结果显示,GD/ScFvMEL抗体的表达量达40%。纯化复性后的GD/ScFvMEL抗体连接量子点制备GD/ScFvMELQDs纳米探针,经扫描电镜、X衍射分析、荧光光谱和SDSPAGE分析证实纳米探针的成功制备。GD/ScFvMELQDs纳米探针能特异性结合A375黑素瘤细胞,不与胃癌细胞MGC803细胞结合。结论：成功制备抗人黑素瘤神经节苷脂单链抗体量子点纳米探针,该探针可特异性结合黑素瘤细胞。     

金纳米笼-量子点-Anti-AFP复合探针对肝癌细胞株的靶向光热治疗

目的 热疗可靶向杀伤乳腺肿瘤细胞,但对肝肿瘤细胞的靶向热疗鲜见报道.本研究探讨金纳米笼-量子点-Anti-AFP复合探针对人肝癌细胞株HepG-2的靶向光热效应.方法 用银立方纳米晶与次氯金酸通过置换反应法制备金纳米笼进行光热治疗,用水热合成法制备CnInS2-ZnS量子点进行细胞荧光显色,将金纳米笼、量子点和Anti-AFP三者耦合成金纳米笼-量子点-Anti-AFP复合探针.采用免疫组化的方法验证肿瘤细胞株是否表达AFP;用免疫荧光法验证复合探针与特定细胞株的靶向结合性;用功率密度为1.5 W/cm2、波长为808 nm激光照射进行光热治疗;用流式细胞术检测光热治疗后的细胞凋亡率;用荧光染色法检测光热治疗后的细胞死亡率.结果 免疫组化实验发现,人肝癌细胞株HepG-2表达AFP,而乳腺癌细胞株Mcf-7不表达AFP.免疫荧光实验表明,复合探针中的Anti-AFP可与HepG-2结合并发出荧光,而不与Mcf-7细胞结合、细胞表面无荧光.光热实验表明,激光照射后,HepG-2细胞的升温明显高于Mcf-7,F=9.369,P<0.001;HepG-2的升温与探针浓度、激光照射时间成正相关、具有一定的时效量效关系.流式细胞术实验表明,光热治疗后,HepG-2的晚期凋亡率明显高于Mcf-7,t=12.551,P<0.001.荧光染色法显示,光热治疗后,95%的HepG-2细胞死亡,而Mcf-7细胞几乎均存活.结论 金纳米笼-量子点-Anti-AFP复合探针可特异性与HepG-2细胞结合,并通过光热效应靶向杀伤HepG-2细胞.     

Exosomes联合卡介苗的体内抗肿瘤效应

目的: 探讨细胞因子诱导杀伤细胞(cytokineinduced killer cell,CIK)对裸鼠胃癌移植瘤的靶向抑制作用。方法:将人胃癌细胞SGC7901注射到裸鼠腹股沟皮下建立胃癌移植瘤裸鼠模型,荷瘤裸鼠随机分为CIK细胞组与成纤维细胞组,分别注射荧光染料SPDiI标记的CIK细胞与成纤维细胞(HFLI)于裸鼠种植瘤对侧腹股沟皮下, 观察其在荷胃癌裸鼠体内各种组织中的分布情况;同时观察CIK治疗后肿瘤的体积大小并计算抑瘤率,病理观察肿瘤的坏死面积。结果: SPDiI标记的CIK细胞注射后10 d主要浓集在荷瘤裸鼠的肿瘤组织,注射局部、肝脏、脾脏和肺脏组织中无CIK细胞或分布极少（P＜001）;标记的成纤维细胞没有出现在肿瘤组织、肝脏、脾脏和肺脏组织,主要集中于注射局部。CIK细胞治疗后裸鼠的移植瘤体积显著小于对照组（P＜0.05）,其抑瘤率为29.82％;移植瘤组织坏死面积评分显著高于对照组（P＜0.01）。结论: CIK细胞对裸鼠胃癌移植瘤有良好的靶向性和杀伤性。     

脂质体介导对反义分子探针肿瘤靶向作用影响的实验研究

[目的]评价脂质体介导对反义分子探针在体外靶细胞摄取以及在荷瘤裸鼠体内特异显像的不同影响情况。[方法]以双功能螫合剂法，制备^99mTc标记的针对人端粒酶催化亚单位（hTERT）mRNA的正、反义探针。分别测定在有／无脂质体介导下．不同处理组的细胞对分子探针的摄取率。建立荷MCF-7乳腺癌裸鼠模型，分别经尾静脉注射有／无脂质体介导的正、反义探针。注射后6h对所有裸鼠进行SPECT显像，分别勾画肿瘤部位与对侧相应部位的感兴趣区，定量分析小同处理组的肿瘤放射性摄取情况。[结果]分子探针的标记率达到76％±5％，放射性化学纯度达到96％以上，比活度达到1850kBq／μg。不论脂质体介导与否，反义探针在不同时相的细胞摄取率均高于正义探针（P〈0．05）；脂质体明显提高细胞对分子探针的摄取，最高可达2．5倍。通过脂质体介导，细胞对反义探针的摄取高峰值提前至2h，并保持稳定水平至3h。荷瘤裸鼠体内显像结果显示：不论脂质体介导与否，反义组的荷瘤裸鼠肿瘤部位可见有明显的放射性浓聚，而正义组的荷瘤裸鼠肿瘤部位未见有明显的放射性浓聚；脂质体介导没有明显增加肿瘤/非肿瘤（T／NT）比值（P〉0．05）。[结论]脂质体在体外能有效地提高分子探针的细胞靶向摄取效率，但是对分子探针在体内的显像效果无明显影响作用。     

减毒沙门氏菌靶向携带共表达质粒p53/siRNA-survivin抗前列腺癌的体内研究

目的：以人减毒沙门氏菌为基因运载体，探讨其携带p53、siRNA-survivin及其共表达质粒p53／siRNA-survivin对肿瘤的靶向性及抗人前列腺癌效应，方法：复制裸鼠前列腺癌皮下移植瘤模型，分别将已构建的重组表达质粒siRNA-scramble，p53，siRNA-survivin及共表达质粒p53／siRNA-survivin转化到减毒人伤寒沙门菌Ty21a，制备成对应重组减毒沙门菌，通过灌胃及瘤内注射到前列腺癌荷瘤裸鼠体内，观察成瘤时间及瘤块大小；以携有绿色荧光蛋白siRNA-survivin的重组茵作为报告基因，流式细胞术检测其在肝、脾、肿瘤组织中靶向分布及基因呈递作用；应用RT-PCR，Western blot，及免疫组织化学染色方法检测治疗后肿瘤组织p53和Survivin基因及蛋白表达。结果：各治疗组均较对照组肿瘤生长缓慢，肿瘤体积小，共表达质粒组与单独基因治疗组比较，肿瘤体积明显缩小，抑瘤率高；荷瘤裸鼠肿瘤组织可测及survivin mRNA表达下降，p53基因表达增强，p53相关基因GRIM-19 mRNA和蛋白表达增强；FCM检测结果发现，减毒人沙门氏菌可在肝、脾及肿瘤组织中存活及表达，但以肿瘤中绿色荧光聚集明显且维持时间较长，其他脏器仅见极少的绿色荧光，结论：人减毒沙门氏菌可作为外源基因载体携带重组质粒，对前列腺癌移植瘤有明显的靶向性，其靶向介导的p53，siRNA-survivin及其表达基因p53／siRNA-survivin，对前列腺癌荷瘤裸鼠有治疗作用，共表达基因较单基因治疗效果好，具有显著的协同作用。     

特异性抗前列腺癌多肽抑制前列腺癌细胞生长的作用研究

背景与目的：本实验组根据PSA具有的肿瘤定位及丝氨酸蛋白酶活性，设计并已获得了包含促细胞凋亡的BH3结构域、抗肿瘤血管形成的VEGF拮抗肽K237和bFGF拮抗肽DG2结构域以及可被PSA特异性水解短肽的复合型多肽APP216的纯化蛋白，并运用细胞培养初步验证了其在体外对前列腺癌细胞的杀伤作用，本文进一步探讨该多肽对人前列腺癌肿瘤异种移植物的杀伤作用，为抗前列腺癌新药的研究奠定基础。方法：利用检测荷瘤裸鼠血清PSA浓度、各组荷瘤裸鼠肿瘤体积、重量变化、各组荷瘤裸鼠治疗前后体重的变化以及病理组织学观察来评估包含有BH3、K237、DG2结构域和能被PSA特异性水解的短肽序列的多肽药物APP216对分泌PSA的前列腺癌细胞系22RV，荷瘤裸鼠及不分泌PSA的前列腺癌细胞系PC3荷瘤裸鼠的肿瘤细胞杀伤作用。结果：前列腺癌细胞22RV，的荷瘤裸鼠治疗后比治疗前血清PSA浓度下降了71．1％、肿瘤体积下降51％，治疗组肿瘤重量比对照组低52％，组织病理学观察镜下可见坏死瘤组织。而前列腺癌细胞PC3的荷瘤裸鼠治疗组肿瘤重量与对照组无明显差别；治疗前后肿瘤体积变化无统计学意义，组织病理学观察癌细胞生长活跃。所有裸鼠重要脏器均未见病理损伤。结论：APP216多肽对分泌PSA的前列腺癌细胞荷瘤裸鼠具有良好的抑制瘤细胞增殖的作用。而对于不分泌PSA的前列腺癌细胞荷瘤裸鼠抑瘤效果不佳。并且该蛋白对其他重要脏器无毒性。     

Cytologic classification of precancer and cancer of the endometrium and esophagus and of malignant lymphomas

The paper discusses cytological classifications of precancer and cancer of the endometrium, esophagus and malignant lymphomas presented by cytologists from five Soviet research institutes of oncology. The classifications were based on the data of 4400 cases in conformity with WHO histologic classifications.     

E-钙粘蛋白及PTEN基因编码蛋白与胃癌浸润转移

目的:观察抑癌基因PTEN蛋白和ECD在胃癌组织中的表达,探讨其与胃癌生物学行为及预后的关系。方法:以兔抗人PTEN多克隆抗体、鼠抗人ECD单克隆抗体,采用SABC免疫组化法,检测100例胃癌手术切除标本中拟测指标的表达。以χ2和Logrank检验对结果做统计学分析。结果:ECD、PTEN蛋白在非癌胃粘膜中均见表达;在胃癌组织中表达下调或缺失。ECD异常表达率为42.0%;弥漫型胃癌异常表达率(48.57%),明显高于肠型胃癌(26.67%),(P<0.05);ECD异常表达与浸润深度有关(P<0.05)。胃癌组织中PTEN蛋白缺失率为59%;弥漫型胃癌缺失率(65.71%)明显高于肠型胃癌(43.33%),(P<0.05);伴淋巴结转移的胃癌缺失率(64.47%)明显高于无淋巴结转移者(41.67%),(P<0.05);PTEN蛋白缺失的患者比阳性表达者预后差(P=0.0066)。65.85%PTEN阳性表达者同时伴ECD正常表达。结论:两种标志物与胃癌浸润转移有关,PTEN表达与胃癌患者预后密切相关。将两种指标联合检测,可作为正确判断胃癌患者预后,指导临床治疗的分子生物学指标。     

Schwannomas of Head and Neck and Review of Literature

Benign nerve cell tumours have been given various names like schwannoma, neurilemmoma, neurinoma, neurofibroma, spindle cell tumours etc. Extra cranial head and neck schwannomas usually present as solitary and well-demarcated lesions. The lesion can cause secondary symptoms, such as nasal obstruction, dysphasia, and hoarseness, depending upon the location of the lesion. Fine needle aspiration cytology, CT scans, and MRI may be of limited help in the diagnosis of schwannomas. The treatment is complete surgical excision of the benign tumour and postoperative histopathological examination establishes the final diagnosis.     

世界卫生组织骨质疏松症防治工作报告和防治建议

引 言 作为对第51号综合处理非传染性疾病预防与控制的世界卫生组织决议的反应,1998年7月WHO成立了致力于不断完善对骨质疏松预防和治疗策略的工作小组。小组成员来自世界各国致力于骨质疏松研究的知名专家。Harry K.Genant为本届主席。这一项世界范围内的骨质疏松教育计划旨在通过世界范围的研究,不断改善对骨质疏松的诊断水平和发展并完善对骨质疏松病人的合理治疗。其重点将以发展中国家为主。并为各国政府及其卫生部门和病人群体提供世界性有关骨质疏松症的总体的、完整的指导性资料。该项研究、教育计划的实施将由世界各国的骨质疏松症研究和治疗机构共同完成,并经权威学术机构、政府和非政府组织进行有针对性的回顾研究,最终由WHO审议通过。     

Prospective study of fruit and vegetable consumption and incidence of colon and rectal cancers

BACKGROUND: Frequent consumption of fruit and vegetables has been associated with a reduced risk of colorectal cancer in many observational studies. METHODS: We prospectively investigated the association between fruit and vegetable consumption and the incidence of colon and rectal cancers in two large cohorts: the Nurses' Health Study (88 764 women) and the Health Professionals' Follow-up Study (47 325 men). Diet was assessed and cumulatively updated in 1980, 1984, 1986, and 1990 among women and in 1986 and 1990 among men. The incidence of cancer of the colon and rectum was ascertained up to June or January of 1996, respectively. Relative risk (RR) estimates were calculated with the use of pooled logistic regression models accounting for various potential confounders. All statistical tests were two-sided. RESULTS: With a follow-up including 1 743 645 person-years and 937 cases of colon cancer, we found little association of colon cancer incidence with fruit and vegetable consumption. For women and men combined, a difference in fruit and vegetable consumption of one additional serving per day was associated with a covariate-adjusted RR of 1.02 (95% confidence interval [CI] = 0.98-1.05). A difference in vegetable consumption of one additional serving per day was associated with an RR of 1.03 (95% CI = 0.97-1.09). Similar results were obtained for women and men considered separately. A difference in fruit consumption of one additional serving per day was associated with a covariate-adjusted RR for colon cancer of 0.96 (95% CI = 0.89-1.03) among women and 1. 08 (95% CI = 1.00-1.16) among men. For rectal cancer (total, 244 cases), a difference in fruit and vegetable consumption of one additional serving per day was associated with an RR of 1.02 (95% CI = 0.95-1.09) in men and women combined. None of these associations was modified by vitamin supplement use or smoking habits. CONCLUSIONS: Although fruits and vegetables may confer protection against some chronic diseases, their frequent consumption does not appear to confer protection from colon or rectal cancer.     

A comparative study of knowledge and awareness of colorectal and breast cancerA comparative study of knowledge and awareness of colorectal and breast cancer

Aims: To assess and compare knowledge and awareness of colorectal cancer and breast cancer in a sample of the general population. Methods: Eleven hundred visitors to six different outpatient clinics, in a University Hospital, were given a study-specific questionnaire, based on educational material from the British Association of Cancer United Patients (CancerBACUP). The questionnaire consisted of 12 statements on the incidence, presentation, detection, treatment and prognosis of colorectal and breast cancer. Results: One thousand and sixty-eight individuals returned the questionnaire. One thousand and four completed questionnaires were analysed. The mean age (SD) of respondents was 50.1 (17.2) years, and the male to female ratio was 2:3. Respondents had read more about breast than about colorectal cancer (60.3%vs 32.4%,P <0.0001, McNemar's test). The proportion of correct answers for each statement on breast cancer was higher than for answers to corresponding items on colorectal cancer. Mean overall scores (95% CI) for breast and colorectal cancer were 88.1 (86.9, 89.2) and 64.4 (62.5, 66.3) respectively, the mean difference (95% CI) being 23.7 (22.0, 25.5). Scores were higher for breast cancer irrespective of age or gender. Conclusion: There is a low level of understanding of colorectal cancer in the general population when compared to breast cancer. This highlights the importance of public education in this common cancer.     

Quality of life and care in leukemia,myeloma and non-malignant disease. Opinions of patients and relatives,and effects of geography and time

In a questionnaire study 140 subjects answered 4200 questions in 1980 and 1986. They consisted of patients with myeloma, acute leukemia, lung carcinoma, and non-malignant disease and their relatives. In 22 additional cases the questionnaire was not answered. The results show that myeloma patients are less content with the general care than leukemia patients (P < 0.05). Similarly, relatives of deceased myeloma patients are less satisfied with the information given to them than relatives of deceased leukemia patients (P < 0.001). The information has improved with time, however, since the patients were more satisfied in 1986 than in 1980 (P < 0.001) and relatives of myeloma patients still alive were more satisfied than relatives of patients who had died earlier (P < 0.001).     

Quality of life and care in leukemia, myeloma and non-malignant disease. Opinions of patients and relatives, and effects of geography and time

In a questionnaire study 140 subjects answered 4200 questions in 1980 and 1986. They consisted of patients with myeloma, acute leukemia, lung carcinoma, and non-malignant disease and their relatives. In 22 additional cases the questionnaire was not answered. The results show that myeloma patients are less content with the general care than leukemia patients (P less than 0.05). Similarly, relatives of decreased myeloma patients are less satisfied with the information given to them than relatives of deceased leukemia patients (P less than 0.001). The information has improved with time, however, since the patients were more satisfied in 1986 than in 1980 (P less than 0.001) and relatives of myeloma patients still alive were more satisfied than relatives of patients who had died earlier (P less than 0.001). The opinions of patients were similar to those of their relatives. However, the relatives of leukemia patients were even more satisfied with the contact with the medical staff than the patients themselves (P less than 0.05). As many as 10-30% of the relatives never gave up hope for their relative's survival. Only two out of 27 deaths were considered not dignified. The lung carcinoma patients reported a less good quality of life (P less than 0.001), and less satisfaction with the information given (P less than 0.01), than the hematological patients from the same year. Similarly, their attitude to the medical care improved less (P less than 0.01), and they were less content with the general care than the leukemia group (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)