婴幼儿咽喉乳头瘤组织人乳头瘤病毒感染的探讨

目的 探讨温州地区人乳头瘤病毒感染和婴幼儿咽喉乳头状瘤的关系。方法 应用聚合酶链反应和核酸斑点杂交技术检测３５例婴幼儿咽喉乳头瘤组织和１０例对照组组织（小儿声带小结）ＨＰＶ６、１１、１６、１８、３３５个型别的ＤＮＡ。结果 乳头瘤组织ＨＰＶ感染率为９１．４％（３０／３５）,其中ＨＰＶ６型检出率为５４．２％（１９／３５）,ＨＰＶ１１型感染率为２５．７％（９／３５）,多重型别ＨＰＶ６＋１１感染率为１１．     

PCR和PCR产物斑点杂交技术检测成人咽喉病变中人乳头状瘤病毒感染

目的 :研究成人咽喉部良、恶性病变与人乳头状瘤病毒 (HPV)感染的关系。方法 :应用聚合酶链反应 (PCR)和斑点杂交技术 ,对 5 5例咽喉不同病变的新鲜组织标本进行 HPV6,1 1 ,1 6,1 8,33共 5型HPV- DNA感染的检测。结果 :在咽乳头状瘤组 HPV感染率为 6 0 % (6 / 1 0 ) ,喉乳头状瘤组为 70 % (7/1 0 ) ,喉鳞状上皮非典型增生组为 2 0 % (1 / 5 ) ,声带息肉组为 2 0 % (1 / 5 ) ,喉癌组为 2 0 % (1 / 5 ) ,声带小结组为 0 (0 / 1 0 )。HPV- DNA型别分布在咽喉良性病变中以 HPV6,1 1 型为主 ,喉癌中以 HPV1 6为主。结论 :本地区成人咽喉良、恶性病变的发生与 HPV感染密切相关。PCR结合核酸斑点杂交法检测 HPV,具有灵敏性高、特异性强的优点 ,值得推广应用。     

人乳头状瘤病毒及其相关基因在中耳胆脂瘤中的表达

目的　探讨人乳头状瘤病毒 (humanpapillomavirus ,HPV)感染在中耳胆脂瘤发生发展中的作用。方法　运用共同引物聚合酶链反应 (polymerasechainreaction ,PCR)和核酸分子斑点杂交法对44例 ( 44耳 )中耳胆脂瘤标本组织中的HPVDNA进行检测 ,并结合其中 35例 ( 35耳 )的病理学检查结果进行对比分析。结果　 12耳 ( 34.3 % )中耳胆脂瘤组织中观察到了HPV感染的损害特征 ;用共同引物PCR法及核酸分子斑点杂交法对 44耳中耳胆脂瘤组织标本进行HPVDNA扩增的阳性率分别为2 9 .5 % ( 13 44 )及 2 5 .0 % ( 11 44 ) ;表现有人乳头瘤病毒损害特征的 12耳中耳胆脂瘤组织HPVDNA检测阳性率为 5 8.3% ( 7 12 ) ,而无此损害特征的 2 3耳中耳胆脂瘤组织HPVDNA检测阳性率为 13.0 %( 3 2 3) ,统计学检验差异有显著性 ( χ2 =7.92 6 ,P <0 .0 0 5 )。结论　HPV感染可能激发中耳胆脂瘤上皮的分裂增殖 ,在中耳胆脂瘤发生发展中起一定的作用 ;侵蚀性乳头瘤样生长和空晕细胞改变可以作为中耳胆脂瘤组织中HPV感染的病理学证据     

人乳头瘤病毒型别与儿童复发性呼吸道乳头状瘤临床病程的相关性

目的　研究人乳头瘤状病毒（human papilloma virus,HPV）型别对儿童复发性呼吸道乳头瘤（juvenile onset recurrent respiratory papillomatosis,JORRP）临床病程的影响。方法　收集38例JORRP患儿新鲜瘤体标本,采用流式荧光杂交法检测HPV型别。将患儿分为HPV6阳性组和HPV11阳性组。量化评分其201例手术,对发病年龄、并发症、临床症状、手术次数、乳头瘤侵袭范围程度等方面进行统计学分析。结果　55.2%（21/38）JORRP患儿感染HPV6,36.8%（14/38）感染HPV11,7.9%（3/38）HPV6/11均阴性。两组患儿在发病年龄、术前临床症状评分、乳头瘤解剖亚区数、Derkay、Dikkers评分方面均有统计学差异。结论　感染HPV11发病年龄更小,侵犯范围更广病灶性状更严重,临床症状更严重,致JORRP临床病程更严重。     

HPV11对小儿喉乳头状瘤预后的影响

目的 :研究人乳头状瘤病毒 (HPV)型别对小儿喉乳头状瘤 (JLP)预后的影响。方法 :应用聚合酶链反应结合斑点杂交技术对 2 5例JLP的石蜡标本进行HPV定型分析 ,并统计HPV11、HPV6 感染组的气管切开率和术后复发率。结果 :HPV总检出率为 96.0 % ,其中HPV11为 5 6.0 % ,HPV6 为 4 0 .0 % ,HPV16、18、33无一例阳性。HPV11感染组的气管切开率为 71.4 % ,术后复发率为 85 .7% ;HPV6 感染组的气管切开率为 3 0 .0 % ,术后复发率为4 0 .0 %。两组分别比较 ,其差异均有显著性意义 (P <0 .0 5 )。结论 :HPV6、11与JLP发生密切相关 ,HPV11感染与JLP的喉梗阻和术后复发率相关 ,HPV11感染可作为JLP预后评判的重要依据。     

鼻腔鼻窦内翻性乳头状瘤临床分期与人乳头状瘤病毒感染的关系

目的 探讨人乳头状瘤病毒(Human Papillomavirus,HPV)与鼻腔鼻窦内翻性乳头状瘤(Nasal Inverted Papilloma,NIP)病变范围的关系.方法 取99例NIP石蜡包埋组织标本采用核酸分子快速导流杂交基因芯片分型技术(HybriMax)对21种HPV亚型同步进行检测和病毒分型,并以24例健康志愿者鼻腔黏膜组织为对照.结果 NIP石蜡标本HPV总感染率64.65%(64/99),对照组均为阴性(0/24).NIP组、对照组比较有统计学意义(P<0.05).NIP不同临床分期间的HPV感染率差异有统计学意义(P<0.05).结论 NIP的发病与HPV感染密切相关,HPV感染能使NIP病变范围扩大,明确NIP中HPV亚型的感染情况,对肿瘤临床行为的预测和指导临床手术有一定意义.     

中耳癌人类乳头状瘤病毒DNA表达的研究

目的 :研究原发性中耳癌人类乳头状瘤病毒 (H PV) DNA的表达情况。方法 :采用多聚酶链反应技术 ,对 5例中耳癌的病理组织蜡块进行 HPV - DNA类型的检测 ,以 8例慢性化脓性中耳炎中耳乳突粘膜为对照组。结果 :5例中耳癌高危型 HPV感染率为 80 % (4 /5 ) ,以 HPV1 6型感染为主 ,而对照组 HPV感染率为 0 % (0 /8) ,经 χ2检验 ,P <0 .0 1。结论 :中耳癌高危型 HPV感染率较高 ,慢性化脓性中耳炎和中耳胆脂瘤以及 HPV感染可能在中耳癌的发生发展中起着重要作用     

人乳头状瘤病毒感染与鼻内翻性乳头状瘤的发病及恶变关系的研究

目的:探讨人乳头状瘤病毒(HPV)感染与鼻内翻性乳头状瘤(NIP)的发病及恶变的关系。方法:将57例NIP患者分为不伴恶变组(45例)和伴恶变组(12例),应用PCR技术,检测其HPV6、11、16、18等4个型别的HPV-DNA,同时以30例鼻息肉患者为对照组。结果:57例NIP患者HPV-DNA总阳性率为64.9%(37/57)。不伴恶变组HPV-DNA阳性率为60%(27/45),均为单一的低危型HPV11型感染;伴恶变组HPV-DNA阳性率为83.3%(10/12),以检出HPV16、18型DNA为主,其中5例为双重感染(4例为HPV16、18型,1例为HPV11、16型),4例为单一型HPV16型感染,1例为单一型HPV11型感染。而30例鼻息肉患者均未检出HPV-DNA。结论:HPV在NIP的发病中有重要作用,而高危型HPV16、18型与NIP恶变可能密切相关。     

儿童咽喉乳头状瘤病毒感染的检测

目的:探讨儿童咽喉乳头状瘤(JOP)与HPV6、HPV11病毒感染及患儿免疫功能的关系。方法:用荧光定量聚合酶链反应(FQ-PCR)检测130例JOP组织病毒DNA,并对其中68例采用流式细胞仪进行细胞免疫功能指标CD3、CD4、CD8检测。结果:130例标本中HPV6和HPV11病毒的检出率为88.46%(115/130),平均拷贝值为5.68±2.65。68例患儿CD3、CD4、CD8百分平均值分别为62.73±8.63、30.54±7.05、26.08±6.93,与对照组比较,差异无统计学意义。结论:FQ-PCR方法简便、准确、特异性强、灵敏度高、定量范围宽,可测定病原体数目为101~1010的不同程度,对JOP病毒的临床诊断、治疗及疗效观察有重要指导作用。     

喉乳头状瘤HPV_(16/18)感染与p53蛋白表达的相关性

目的 :了解喉乳头状瘤组织内 HPV1 6 / 1 8的感染与抑癌基因 p5 3变异的关系 ,以及 HPV感染在喉乳头状瘤发病中的作用。方法 :采用 PCR和免疫组化技术 ,检测 35例喉乳头状瘤组织中 HPV1 6 / 1 8DNA及 p5 3蛋白的表达。结果 :2 4例组织中检出 HPV1 6 / 1 8DNA(6 8.6 % ) ;19例 p5 3蛋白呈过度表达 (5 4.3% ) ;在 12例中同时检出HPV1 6 / 1 8DNA和 p5 3蛋白过度表达 (34 .3% )。结论 :提示 HPV1 6 / 1 8感染和 p5 3变异与喉乳头状瘤的发生明确相关 ,其内在分子机理及临床意义有待进一步阐明。     

小儿喉乳头状瘤HPV-DNA及体液免疫检测

目的：探讨小儿喉乳头状瘤（ＪＬＰ）人乳头瘤病毒（ＨＰＶ）感染途径及发病机理。方法：采用ＰＣＲ及ＰＣＲ产物斑点杂交技术检测ＪＬＰ组织ＨＰＶ－ＤＮＡ;散射免疫比浊法测定血清Ｉｇ及补体Ｃ３。结果：ＪＬＰ组织ＨＰＶ总感染率为９５％（１９／２０）,其中ＨＰＶ。型为５５％（１１／２０）,ＨＰＶ１１为３０％（６／２０）,ＨＰＶ６＋１１型为１０％（２／２０）;ＪＬＰ患者血清ＩｇＧ、ＩｇＡ、ＩｇＭ、Ｃ３值正常,对照     

Detection of human papillomavirus genome in nasolaryngeal papillomas using digoxigenin labeled DNA probes

It is being reported that human papillomavirus (HPV) has been implicated in the pathogenesis of various neoplastic lesions of the genital organs. To investigate the etiological role of HPV and its types in nasolaryngeal papillomas, we retrospectively analyzed HPV genomes by nucleic acid hybridization methods; for detecting DNA and mRNA, we employed the recently developed nonradioactive (digoxigenin labeled) DNA probes and compared the results by radioisotope methods. In total, 43 cases of papillomatous lesions were examined. They were verruca vulgaris of the nasal vestibule (Nr = 2), nasal inverted papilloma (IP, Nr = 26), and laryngeal papilloma (Nr = 15). HPV types examined were type 2, 6, 11, 16 and 18. Two cases of verruca vulgaris were shown to contain HPV-2 DNA and its mRNA by in situ hybridization. HPV-11 DNA was detected in 3 cases (12%) of nasal inverted papilloma whereas HPV-16 was detected in 1 case (4%); the latter case was associated with squamous cell carcinoma. These results suggest that HPV may be implicated in the development of IP, and HPV-16 may play an important role in the malignant transformation of IP. In the cases of multiple laryngeal papilloma (Nr = 8, one juvenile type and 7 adult type), either HPV-6 or HPV-11 was detected at the high rate (6/8, 75%). The presence of the HPV genomes provides strong evidence for the HPV etiology of these laryngeal papillomas. Whereas in the cases of adult single laryngeal papilloma (Nr = 7), HPV was not detected. Technically, the sensitivity of digoxigenin (DIG) labeled DNA probe was almost same as 35S labeled probe by dot blot hybridization, thus we applied DIG labeled probe to Southern blot hybridization with low background. By in situ hybridization using digoxigenin labeled probes, the rates of HPV detection were almost equal to those by 35S labeled probes.(ABSTRACT TRUNCATED AT 250 WORDS)     

新疆部分地区儿童复发性喉乳头状瘤HPV6和HPV11病毒感染在维吾尔族及汉族患者之间的差异

目的：研究新疆部分地区喉乳头状瘤病毒HPV6,HPV11在汉族及维吾尔族儿童复发性喉乳头状瘤（JRRP）患儿中的表达差异。方法：采用聚合酶链反应（PCR）技术对喉乳头状瘤组织中HPV11和HPV6 DNA进行定型分析,结合回顾性分析1996—01—2008—03期间在新疆医科大学第一附属医院耳鼻咽喉科连续收治的42例JRRP患者。结果：HPV6／11阳性检出率97．61％（41／42）,HPV6阳性36．58％（41／15）,HPV11阳性63．41％（41／26）,HPV6阳性组中维族HPV6阳性53．33％（8／15）,汉族HPV6阳性46．67％（7／15）,HPV11阳性组中维族HPVll阳性65．38％（17／26）,汉族HPV11阳性34．61％（9／26）。结论：新疆部分地区JRRP以HPV11,6感染为主,HPV11感染者占多数,HPV病毒类型与维汉间的发病率之间差异无统计学意义。     

Detection and typing of papillomavirus DNA in formaldehyde-fixed paraffin-embedded tissue

Clinical specimens from nine patients with papillomatosis of the vocal cords and three patients with vocal cord polyps were evaluated for the presence of human papillomavirus (HPV) DNA using two complementary molecular hybridization techniques. In one method, involving polymerase chain reaction (PCR) amplification, HPV DNA sequences were replicated in vitro from tissue DNA extracted from paraffin sections prior to hybridization. Polymerase chain reaction amplification was compared with the standard method of Southern blot hybridization. Results of the two techniques for all nine laryngeal papillomas agreed completely: five patients harbored HPV type 6 and four HPV type 11. Both PCR amplification and Southern blot hybridization found two of the three polyps to be free of HPV infection, while PCR detected HPV type 18 in one polyp specimen that was reported negative by Southern blot hybridization, suggesting a greater sensitivity of PCR. Our results demonstrate that PCR amplification is as reliable and at least as sensitive as Southern blot hybridization. Moreover the PCR technique opens the way to the undertaking of a whole variety of retrospective studies using formaldehyde-fixed paraffin-embedded tissues.     

Human papilloma virus infection and expression of p16 protein in laryngeal papilloma and laryngeal carcinoma]

OBJECTIVE: To evaluate the role of human papilloma virus (HPV) infection and inactivation of p16 gene in laryngeal papilloma (LP) and laryngeal squamous cell carcinoma (LC). METHODS: HPV consensus primers direct in situ polymerase chain reaction (ISPCR) and immunohistochemical method were applied to detect the presence of HPV genomes (1, 6, 8, 11, 13, 16, 18, 30, 31, 32, 33, 45, 51) and the expression of p16 protein respectively in 93 cases of formalin-fixed, paraffin-imbedded specimens, which contained 46 cases of LPs [adult-onset laryngeal papilloma (ALP) 21, juvenile-onset laryngeal papilloma (JLP)25], 26 cases of LCs, 6 cases of normal tissues adjacent to carcinoma, and 15 cases of vocal noduli. RESULTS: (1) The difference of positive rates of HPV-DNA in JLP group (84%, 21/25) and other groups were statistically significant (chi 2 test, P < 0.05). The difference of positive rates of HPV-DNA in ALPs(38.1%, 8/21), in LCs(19.2%, 5/26), in vocal noduli(0%, 0/15), and in normal tissues adjacent to carcinoma(0%, 0/6) were not significant statistically (chi 2 test or Fisher's exact probability test, P > 0.05). (2) The positive rates of expression of p16 protein in ALP group(57.1%, 12/21) and LC group(38.5%, 10/26) were significantly lower than that in vocal nodule group(93.3%, 14/15), in JLP group(88%, 22/25), and in normal tissues adjacent to carcinoma group (100%, 6/6) (chi 2 test or Fisher's exact probability test, P > 0.05). There were no significant differences of positive rates of expression of p16 protein between ALP group and LC group, and between JLP group and vocal nodule group (chi 2 test, P > 0.05). (3) In LPs, the difference of positive rates of p16 protein expression between HPV positive cases and HPV negative cases was significant statistically (chi 2 test, P < 0.05). In LCs, there was no difference in p16 protein expression rate between the two teams(Fisher exact probability test, P > 0.05). CONCLUSION: The pathogenesis of JLP is closely associated with HPV infection and not associated with the inactivation of p16 gene. Conversely, the pathogenesis of ALP and LC is associated with the inactivation of p16 gene and not associated with the HPV infection.     

人咽喉部良恶性肿瘤与乳头状瘤病毒关系的研究

采用免疫组化及ＤＮＡ斑点杂交技术检测人咽喉部乳头状瘤及鳞状细胞癌组织中人乳头状瘤病毒（ＨＰＶ）壳蛋白抗原及ＨＰＶ６、１１、１６、１８型ＤＮＡ。１１例乳头状瘤ＨＰＶ抗原与ＨＰＶ ＤＮＡ阳性率均为４５．５％。２２例鳞状细胞癌ＨＰＶ抗原阳性率２２．７％，ＨＰＶ ＤＮＡ阳性率２７．３％。乳头状瘤ＨＰＶ检出率与组织学检查的结果相符。提示咽喉部乳头状瘤及鳞状细胞癌的发生、发展与ＨＰＶ感染有关。     

Human papillomavirus in laryngeal papillomas and in adjacent normal epithelium

Eleven adults with laryngeal papillomas were studied for the presence of human papillomavirus (HPV) DNA by in situ hybridization. As well as from the papillomas, three additional biopsies were taken from the normal-appearing mucosa as follows: the involved vocal cord, the opposite vocal cord (when the papilloma was unilateral), and from the ventricular fold on the side of the lesion. These normal tissues were analysed by polymerase chain reaction (PCR) to detect HPV DNA. All except one of the 11 papillomas contained HPV DNA; nine were HPV 6/11 DNA positive and one positive for HPV 16 DNA. The normal-appearing laryngeal mucosa harboured HPV DNA in eight out of 11 patients. The present results strongly support the concept that the adult-type laryngeal papilloma is an HPV-induced lesion, mostly due to HPV types 6 and 11. The persistence of HPV DNA in the adjacent normal epithelium is consistent with the frequent recurrence of these lesions.     

原位杂交在成年和幼年型喉乳头状瘤HPV病因学的对比研究

探讨成年和幼年型喉乳头状瘤ＨＰＶ感染发病差异及其影响因素。方法：用地高辛配基（Ｄｉｇｏｘｉｇｅｎｉｎ）标记ＨＰＶ６和ＨＰＶ１１型作探针，原位核酸杂交方法在２９例成年型喉乳头状瘤（ＡＬＰ）和２１例幼年型喉乳头状瘤（ＪＬＰ）石蜡包埋标本检测ＨＰＶ同源序列。结果：ＡＬＰＨＰＶ６和ＨＰＶ１１阳性率分别为４１４％（１２／２９）和４８３％（１４／２９）；ＪＬＰＨＰＶ６及ＨＰＶ１１阳性率均为７６２％（１６／２１）。ｘ２统计示：两型喉乳头状瘤ＨＰＶ６及ＨＰＶ１１阳性率明显不同（ＨＰＶ６ｘ２＝５９９，ＨＰＶ１１ｘ２＝３９５，Ｐ均小于００５）。结论：１）ＡＬＰ和ＪＬＰＨＰＶ感染发病存在差异。２）ＡＬＰ除了ＨＰＶ感染外，其促发因素不可忽视，ＪＬＰ更倾向于依赖ＨＰＶ感染而发病。