新疆汉族和维吾尔族鼻咽癌组织的PCR和免疫组化检测比较

目的：了解新疆汉族维吾尔族（维）族鼻咽癌（ＮＰＣ）发病因素的差别。方法：用ＰＣＲ与免疫组化技术对７３例ＮＰＣ组织（汉族４１例,维族３２例）分别进行ＥＢ病毒脱氧核糖核酸（ＥＢＶ－ＤＮＡ）、ＥＢ病毒核抗原（ＥＢＮＡ２）、潜伏膜蛋白（ＬＭＰ－１）检测。     

鼻咽癌放疗后EB病毒相关抗体的调查

目的 调查放疗后鼻咽癌患者人群EB病毒相关抗体VCA-Ig A或NA1-Ig A的变化趋势。方法 114例放疗后鼻咽癌患者,回顾性调查分析其EB病毒相关抗体VCA-Ig A或NA1-Ig A的变化趋势,并与健康体检人群56877人进行比较,分析鼻咽癌患者放疗后不同时期的EB病毒相关抗体变化特征及其演变趋势。结果 结果显示,放疗后鼻咽癌患者EB病毒相关抗体VCA-Ig A阳性率48.25%,NA1-Ig A阳性率25.42%;健康对照组人群VCA-Ig A阳性率5.74%,NA1-Ig A阳性率2.45%,组间差异具有显著性统计学意义（P<0.01）。鼻咽癌患者治疗后5年内阳性率最高,20年以上者阳性率最低,统计学意义不显著（P>0.05）。结论 鼻咽癌患者放疗后EB病毒相关抗体阳性率仍明显高于健康人群,需要加强监测。     

血浆EB病毒DNA含量定量检测对鼻咽癌的诊断和预后评价

目的探讨血浆EB病毒DNA定量检测对鼻咽癌的诊断和预后评估价值.方法收集2002年2月～2003年6月在本院确诊的初诊鼻咽癌患者100例,放疗后鼻咽癌患者106例,以及健康对照者40例,分别采集血浆标本,应用Real-time PCR方法检测EB病毒DNA含量,比较健康者和初诊鼻咽癌患者、放疗后转移或复发及持续临床缓解鼻咽癌患者的血浆EB病毒DNA拷贝数差异.结果初诊鼻咽癌患者、放疗后转移和复发鼻咽癌患者的血浆中游离EB病毒DNA检出阳性率分别为90.7%（97/100）,96.5%（27/28）和100.0%（18/18）,明显高于健康对照组的7.5%（3/40）,持续临床缓解鼻咽癌患者的10.0%（6/60）和其他肿瘤患者的阳性检出率（P＜0.001）.血浆EB病毒DNA定量检测对初诊鼻咽癌患者的诊断敏感性为97.0%,特异性为92.0%.另有3例持续临床缓解而血浆EB病毒DNA水平升高的鼻咽癌患者,在随后的6个月随访中,最终出现肿瘤复发或转移.结论血浆EB病毒DNA定量检测是一种敏感而可靠的实验方法,对鼻咽癌的诊断、病情监测和预后评估具有重要价值.     

MDM 2基因在鼻咽癌中的表达及其与p53蛋白表达、EB病毒潜伏感染的关系

目的：探讨ＭＤＭ２基因在鼻咽癌（ＮＰＣ）中的表达情况及其与ｐ５３蛋白表达、ＥＢ病毒（ＥＢＶ）潜伏感染的关系。方法：运用非同位素原位杂交、免疫组化和多聚酶链反应技术对４６例ＮＰＣ、１２例慢性鼻咽炎症粘膜（ＣＩＮＥ）分别进行ＭＤＭ２ｍＲＮＡ、ＭＤＭ２蛋白、ｐ５３蛋白及ＥＢＶ－ＤＮＡ的检测。结果：４６例ＮＰＣ中,１４例ｍＲＮＡ及ＭＤＭ２蛋白高表达,３８例为ｐ５３蛋白阳性,４３例为ＥＢＶ－ＤＮＡ阳性,１２     

EB病毒Rta蛋白抗体检测对鼻咽癌病理分级的临床价值

目的探讨EB病毒Rta蛋白抗体对鼻咽癌病理分级的临床价值。方法选择我院诊治的鼻咽癌患者共计68例,选取同一时期来我院体检健康人54例,测定血清Rta-IgG水平,血清Rta-IgG与鼻咽癌病理分级采用spearman相关分析。结果7观察组(I期、II期、III期、IV期)血清Rta-IgG水平均高于对照组,鼻咽癌患者中I期、II期、III期、IV期血清Rta-IgG水平依次升高,有差异(P<0.05)。血清Rta-IgG水平与鼻咽癌病理分级之间呈正相关(P<0.05)。结论EB病毒Rta蛋白抗体对鼻咽癌病理分级有预测价值。     

鼻咽癌患者唾液和血清及外周血白细胞中EBV-DNA的定量检测

目的对鼻咽癌患者唾液、血清及外周血白细胞中的EBV-DNA进行检测,分析唾液、血清、外周血白细胞中EB病毒的DNA含量与鼻咽癌的相关性。方法对40例初诊鼻咽癌患者和50例正常人的唾液、血清及外周血白细胞中的EBV-DNA进行荧光聚合酶链反应(FQ-PCR)检测。结果EBV-DNA在鼻咽癌患者血清中的检出率为67.5%,显著高于正常人血清中的EBV-DNA检出率2%(χ2=44.48,P<0.001);鼻咽癌患者的唾液、外周血白细胞EBV-DNA检出率与正常人相比,无显著性差异;鼻咽癌患者和正常人的唾液EBV-DNA拷贝数与外周血白细胞EBV-DNA拷贝数呈正相关性;鼻咽癌患者血清EBV-DNA拷贝数与TNM分级有关,分级越晚含量越高。结论 EB病毒长期存在于多数人的唾液中,也可存在于外周血B淋巴细胞中。血清EBV-DNA检测在鼻咽癌的临床诊断中具有较高的敏感性和特异性,有助于鼻咽癌的早期诊断;血清EBV-DNA拷贝数与TNM分级有关,可在分子水平对鼻咽癌的TNM分级进行补充。     

EBV潜伏膜蛋白-1表达与鼻咽癌增殖和凋亡关系的研究

目的:研究鼻咽癌(nasopharyngeal carcinoma,NPC)组织中EBV基因EBER-1和LMP1的表达,凋亡相关基因Bcl-2和Bax,Fas和Fas-L,Caspase 3表达及其与p53蛋白的积聚、肿瘤细胞增殖和凋亡的关系。方法:分别用免疫组化染色、原位杂交和凋亡原位标记(TUNEL)技术研究NPC组织中EBV表达、凋亡相关基因表达、肿瘤增殖活性和凋亡活性。结果:87例NPC中,EBER1原位杂交(ISH)检出率为100％,EBV-LMP1阳性率63.2％(55/87),p53蛋白阳性率95.4％(83/87),肿瘤细胞Ki67核阳性率96.6％(84/87),鼻咽癌p53阳性强度与EBV-LMP1表达呈显著正相关(P<0.0005)。本组NPC病例Bcl-2和Bax阳性率分别为69.0％(60/87)和6.2％(57/87),Fas和Fas-L阳性率分别为87.2％(75/86)和54.7％(47/86),Caspase 3阳性率为62.1％(54/87)。NPC组织 Bcl-2的表达与Ki67的表达明显负相关(P<0.005),肿瘤细胞的增殖活性(Ki67的表达)与p53蛋白积聚显著正相关(P<0.0005)。Caspase 3表达与EBV-LMP1的表达呈负相关(P<0.001),但与Fas-L的表达呈正相关(P相似文献   

鼻咽癌患者血清唾液酸动态观察

目的探讨血清唾液酸(sialicacid,SA)在鼻咽癌的诊断价值及作为病情动态监测指标的可行性。方法对50例鼻咽癌患者治疗前、治疗结束时、治疗后2年的血清唾液酸进行检测,并与EB病毒衣壳抗原免疫球蛋白A(Epstein-Barrvirus-viralcoatantigensIgA,EBVVCA-IgA)作比较;以健康成年人、头颈部良性病变者各50例,作为对照。结果鼻咽癌患者治疗前血清唾液酸和EBV-VCA-IgA阳性率均明显高于健康人,差异有统计学意义(P值均<0.01);不同时期血清唾液酸与EBVVCAIgA平均值呈明显正相关。血清唾液酸在鼻咽癌患者治疗前、后及随访期变化差异有统计学意义(P<0.01),治疗前阳性率达94.0%(47/50),治疗结束时阳性率2.0%(1/50),随访期有复发者阳性率96.2%(25/26),未复发者阳性率4.2%(1/24);而EBVVCAIgA变化不太明显。作为诊断试验,血清唾液酸对鼻咽癌的敏感度为94.0%,高于EBVVCAIgA的90.0%;特异性为93.0%,低于EBV-VCA-IgA的96.0%。结论血清唾液酸可能成为鼻咽癌病情变化的动态监测指标,结合EBV-VCA-IgA的检测,可用于高危人群的普查、筛选,以及对临床疗效的判断和预后的评估。     

鼻咽癌相关的癌基因研究进展

鼻咽癌是头颈部的恶性肿瘤，最常见于我国和东南亚国家，我国广东、广西、湖南、福建、江西等地是鼻咽癌的高发区。黄种人患鼻咽癌多见，而白种人和欧美国家患此病者较为罕见。中国人的鼻咽癌发病率是印度人的30倍，是13本人的60倍，其病死率占全部恶性肿瘤病死率的第8位(男)和第9位(女)。鼻咽癌的发生与多种因素有关，目前认为比较明确的有3方面因素：遗传因素、环境致癌因素以及EB病毒感染。     

鼻咽癌患者放疗后引起感音神经性聋的临床观察

目的探讨鼻咽癌患者放疗后不同时期感音神经性聋的程度及发生率。方法选择无分泌性中耳炎的鼻咽癌患者28例，利用纯音测听和听性脑干反应（ABR），结合声导抗和耳声发射测试患者放疗前和放疗后1个月、1年、2年和5年的纯音听阈及波Ⅰ、Ⅲ、Ⅴ潜伏期、Ⅰ～Ⅴ波间期、鼓室压和镫骨肌声反射，以及部分受放射耳的诱发性耳声发射。结果语频（0．5～4．0kHz）和高频（8kHz）平均骨导听阈情况：①放疗后1个月较治疗前分别提高7．1dB和25．7dB，与放疗前比较差异有统计学意义（P〈0．001）；②放疗后1年分别提高17．6dB和28．1dB，与放疗前比较差异有统计学意义（P〈0．001），与放疗后1个月比较差异也有统计学意义（P值均〈0．001或P〈0．05）；③放疗后2年分别提高21dB和27．4dB，与放疗后1年比较差异仅语频有统计学意义（P〈0．05）；④放疗后5年分别提高26．7dB和35．8dB，与放疗前、放疗后1个月、1年和2年比较差异均有统计学意义（P值均〈0．001）。其中语频和高频听力损害大于15dB的发生率分别为37．5％～94．7％和85．4％～97．4％；听力损害大于30dB的发生率分别为14．6％～63．2％和37．5％～73．7％。平均ABR波Ⅰ、Ⅲ和Ⅴ潜伏期及Ⅰ～Ⅴ波间期的情况：放疗后1个月与放疗前比较无明显延长（P〉0．05）；放疗后1年和2年，均较放疗前和放疗后1个月明显延长，差异有统计学意义（P〈0．05），1年和2年组比较差异无统计学意义（P〉0．05）；放疗后5年较放疗前显著延长（P〈0．001），与放疗后1年和2年比较，波Ⅰ、Ⅲ和Ⅴ潜伏期明显延长（P〈0．05），而Ⅰ～Ⅴ波间期无明显延长（P〉0．05）。放疗1年后10耳中有7耳，放疗5年后7耳中有4耳诱发性耳声发射正常，但ABR均明显异常。结论放射所致的感音神经性聋可发生在部分患者放疗后的早期，特别是高频；随放疗后时间延长，听力损害的发生率增加，程度加重；损害可发生在耳蜗或（和）蜗后听觉通路，表明听觉系统不同部位和不同个体对放射损伤的敏感性可能存在差异。     

Morphometric and stereologic nuclear values obtained from the study of a population suffering from nasopharyngeal carcinoma

Computer nuclear morphometry and stereology are attractive methods because its objectivity and cheapness allowing histologic diagnosis when identifying minimal variations respectively the normality and also detect negligible disparities between anormal cells which could escape to the assessment of the pathologist. We present the data gained from several morphogenic and stereologic parameters resulting of measurements of tumoral cells procured from 40 patients with nasopharyngeal carcinomata. Middle values have been: nuclear area 27.70 microns 2; nuclear perimeter 20.80 microns; nuclear factor of form 0.81 microns; nuclear outline index 4.01; nuclear orientation angle 87.29 degrees; nuclear ellipsiticity 704.14; nuclear regularity 61.83; middle lineal length 4.30, middle linear distance 107.94; and nuclear volume 118.80 microns 3. Our series is the largest studied till now of all found in the literature. Comparison our data with those of previous publications.     

鼻咽癌患者鼻咽组织和血清中EBV检测

目的：比较血清爱泼斯坦－巴尔病毒（ＥＢＶ）抗体滴定度测定与癌组织ＥＢＶ基因组检测对鼻咽癌（ＮＰＣ）的诊断价值。方法：１４６例患者采用双盲法测定血清ＥＢＶ－ＶＣＡ－ＩｇＡ,ＥＢＶ－ＥＡ－ＩｇＡ和活检组织ＥＢＶ－ＤＮＡ（ＰＣＲ）。全组病例按活检病理检查结果分析：ＮＰＣ组,非ＮＰＣ组（对照组）。结果：ＮＰＣ组中ＥＢＶ－ＤＮＡ（ＰＣＲ）,ＥＢＶ－ＶＣＡ－ＩｇＡ和ＥＢＶ－ＥＡ－ＩｇＡ的阳性率分别为９０．８％     

CK13基因与EB病毒DNA在鼻咽癌中的表达及临床意义

目的 探讨细胞角蛋白基因13（CK13）和EB病毒DNA在人鼻咽癌（NPC）组织中的表达及意义。方法 应用Northern杂交对32例NPC和8例慢性鼻咽炎（CINE）组织进行CK13mRNA水平检测,同时应用PCR方法进行EB病毒DNA检测。结果 ＣＫ１３基因在鼻咽炎组织中高表达,在ＮＰＣ组织中的表达显著下调。下调的频率达６５.63%(21/32)。结论 ＮＰＣ组织的分化可能与ＣＫ１３基因的表达下调有关。     

Epstein-Barr virus DNA measured in nasopharyngeal brushings in patients with nasopharyngeal carcinoma: pilot study

OBJECTIVE: We measured the amount of tumour-derived Epstein-Barr virus (EBV) deoxyribonucleic acid (DNA) in the nasal brushings of nasopharyngeal carcinoma (NPC) patients to determine the correlation with tumour load and response to treatment. MATERIALS AND METHODS: Twenty-eight patients with NPC were included in the study. Baseline measurements of EBV from nasopharyngeal brushings were obtained from 26 patients prior to treatment. A follow-up sample was available from 11 of these patients post-treatment and from 2 additional patients who did not have a baseline sample. Quantitative real-time polymerase chain reaction (PCR) using SYBR Green I fluorescent dye was used to detect the EBV DNA copy number. RESULTS: Nasopharyngeal brush biopsies showed a high copy number of EBV DNA in most of the pretreatment samples (median 9714 copies/mL). The highest copy number detected was 14536944 copies/mL in one sample. In the post-treatment follow-up samples, the copy number was significantly lower (median 6 copies/mL). CONCLUSIONS: We have demonstrated that EBV DNA can be detected in the brush biopsies from NPC patients using quantitative real-time PCR. These pilot data suggest that nasopharyngeal brushings with PCR detection of EBV may be an effective tool for determining local tumour response. The potential of this technique as an NPC tumour marker for post-treatment follow-up is being validated with larger patient numbers.     

A basic study on the association of Epstein-Barr virus with nasopharyngeal carcinoma--detection and genotypic analysis of Epstein-Barr virus associated with nasopharyngeal carcinoma in Japanese patients]

Epstein-Barr virus (EBV) is known to be associated with two human malignant diseases, nasopharyngeal carcinoma (NPC) and endemic Burkitt's lymphoma. In this study, the genotypes of EBV in tissues from 13 NPC patients in Japan were analyzed by Southern blot hybridization using EBV genome fragment probes. Ten of the cases contained reiterated sequences (EBV BamHI-H, -B1*, -K fragments), showing that only one genotype was detected in each specimen. One of these had a BamHI fragment containing a fused sequence of BamHI-Y and -H. In all except one case, a single-sized EBV-joined terminus was observed in each NPC specimen, implying evolution of the carcinoma from a single EBV-infected cell. One metastatic lymph node (which was not a primary epipharyngeal tumor) contained EBV with heterogeneous termini suggesting production of linear virion DNA. The type C variant resulting from loss of a BamHI site between the BamHI-W1* and -I1* regions was observed in 7 of the 10 cases, and the other 3 cases had a separated BamHI-I1* fragment. As reported by Lung et al. (Virology, 177: 44-53, 1990), the type C variant appears to be dominant among Japanese strains, as it is in Southern China. In contrast to their findings, however, the "f" variant with an extra BamHI site in the BamHI-F region which they found to be strongly associated with NPC specimens from Southern China, was detected in only one case. The present study, therefore, did not support the specific association of the "f" variant with NPC in Japanese patients. We conclude that the EBV in NPC tissues exists in variants. Further studies along these lines, could help to explain the epidemiology of EBV.     

晚期鼻咽癌同步放化疗患者营养状况监测及院内感染分析

目的：观察鼻咽癌同步放化疗患者治疗前后的营养状况和院内感染情况,探讨预防感染的有效措施。方法：对82例鼻咽癌接受同步放化疗的患者治疗前后的营养状况以及院内感染和治疗情况进行回顾性分析。结果：同步放化疗前后患者营养状况,包括体重、血红蛋白及血清白蛋白之间差异有统计学意义。82例患者中发生院内感染者63例（76．83％）;口鼻、咽部黏膜感染者76例（92．68％）;黏膜感染伴有皮肤带状疱疹者2例（2．44％）;肺部感染者3例（3．66％）;败血症1例（1．22％）。共分离病原菌39株,病原菌其中革兰阴性菌最为多见,共23株（58．97％）;其次真菌12株（30．77％）;革兰阳性菌3株（7．69％）;带状疱疹病毒1株（2．56％）。结论：晚期鼻咽癌同步放化疗患者治疗期间应做好营养监测和营养支持,加强饮食指导和干预以减少其不良反应的发生,提高患者的躯体耐受性及生活质量。同期放化疗期间院内感染以口咽黏膜感染革兰阴性菌发生率最高,应重视咽拭子培养,必须加强口腔护理并合理用药。     

鼻咽癌患者外周血γδT细胞对鼻咽癌细胞的细胞毒活性测定

T细胞受体(T cell receptor，TCR)γδ表型细胞与TCRαβ细胞不同，识别抗原的方式类似于免疫球蛋白，以主要组织相容性复合体非限制性方式识别抗原分子，无需抗原呈递分子配合，直接识别广谱肿瘤抗原。我们以抗体固相法体外扩增7例鼻咽癌(nasopharyngeal carcinoma，NPC)患者及6例健康人外周血的γδT细胞，经原代NPC细胞同步培养，得到高纯度的NPC细胞。流式细胞仪检测γδT细胞纯度，研究两种来源的γδT细胞对Daudi、CNE1、CNE2和新建的传代培养NPC细胞的细胞毒作用，探讨NPC患者外周血中γδT细胞的抗肿瘤作用。     

Benign nasopharyngeal masses and human immunodeficiency virus infection

Manifestations of the acquired immunodeficiency syndrome are common in the head and neck and are becoming well known to the otolaryngologist. We present a series of seven patients who complained of nasal obstruction and hearing loss and were found, on examination, to have large obstructing nasopharyngeal masses and otitis media with effusion. Biopsy revealed benign lymphoid proliferation. Because of a suspicion of human immunodeficiency virus infection by history, antibody titers were obtained and were found to be positive in all cases. With the known increased rate of aggressive extranodal B-cell lymphomas in human immunodeficiency virus-infected patients, its existence in the nasopharynx should be ruled out histologically in symptomatic patients. Nasal obstruction and hearing loss secondary to nasopharyngeal lymphoid proliferation in high-risk patients can be an early sign of human immunodeficiency virus infection. Patients presenting with this clinical entity should be advised to have serologic testing and further treatment and counseling if necessary.