喉乳头状瘤患儿血清人乳头状瘤病毒11b病毒样颗粒抗体检测及意义

目的　制备人乳头状瘤病毒 (humanpapillomavirus,HPV) 11b型病毒样颗粒 (viruslikeparticles ,VLP)检测儿童喉乳头状瘤 (juvenilelarynxpapilloma ,JLP)患者血清抗HPV11bVLP抗体 ,探讨与疾病发生、发展的关系。方法　采用重组杆状病毒昆虫细胞系统制备HPV11bVLP。通过酶联免疫吸附试验 (enzymelinkedimmunosorbentassay,ELISA)方法检测 4 6份复发性儿童喉乳头状瘤患儿不同发病时期的血清和 2 0份健康对照组的血清抗体。将标本分 5组 :A组 :健康对照 (2 0份 ) ;B组 :发病 1年(15份 ) ;C组 :发病 2年 (15份 ) ;D组 :未复发 1年 (8份 ) ;E组 :未复发 2年 (8份 )。结果A、B、C、D、E各组血清抗体吸光度 (A)值平均 ( x±s)分别为 0 0 73± 0 0 35 ;0 12 0± 0 0 4 9;0 137± 0 0 5 7;0 5 18±0 12 2 ;0 5 5 7± 0 14 4 ,B、C、D、E各组与A组比较差异均有显著性意义 (P <0 0 5 )。未复发组 (D +F组 )和复发组 (B +C组 )间A均值比较差异有高度显著性意义 (t=14 90 ,P <0 0 0 0 1)。两组年龄比较差异有显著性意义 (6 2 5 0± 1 6 93和 3 2 2 7± 1 4 5 3;t=6 76 7,P <0 0 0 0 1)。结论　HPV感染的JLP患者血清能产生抗HPV抗体 ,疾病发展和预后与血清抗病毒中和抗体水平密切相关。HPV11bVLP及血     

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。     

喉癌和喉乳头状瘤组织中人乳头状瘤病毒和p16蛋白的检测

目的探讨人类乳头状瘤病毒（humanpapillomavirus,HPV）感染和抑癌基因p16的失活与喉癌和喉乳头状瘤（laryngealpapilloma,LP）发生的相关性,以进一步阐明喉癌和LP的病因和发病机理。方法收集LP46例,其中成人型喉乳头状瘤（adult-onsetLP,ALP）21例,青少年型喉乳头状瘤（juvenile-onsetLP,JLP）25例、喉癌26例、癌旁正常组织6例、声带小结15例,用标记的HPV1,6,8,11,13,16,18,30,31,32,33,45,51通用引物直接法原位聚合酶链反应（polymerasechainreaction,PCR）方法和免疫组化（SP法）方法分别检测HPV-DNA和p16蛋白。结果①HPV阳性率JLP组（84%,21/25）显著高于ALP组（38.1%,8/21）、喉癌组（19.2%,5/26）、声带小结组（0/15）和癌旁组织组（0/6）（χ     

儿童喉乳头状瘤研究进展[综述]

喉乳头状瘤是儿童最常见的呼吸道良性肿瘤,主要临床表现为声嘶、喘鸣和呼吸困难,严重者可出现完全性喉梗阻导致窒息。儿童喉乳头状瘤的病原体主要为人乳头状瘤病毒（human papilloma virus,HPV）,且主要为低危型HPV6、11型,HPV11型感染和发病年龄早更容易导致不良预后。目前认为幼儿HPV感染途径主要为宫内感染、经阴道接触感染,但母体感染HPV并不是终止妊娠和剖宫产指征。手术是目前主要的治疗方式,疫苗接种等辅助治疗日益发挥作用。     

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。     

儿童喉乳头状瘤与人乳头状瘤病毒的相关性及其免疫综合治疗

对2002～2005年收治的60例儿童喉乳头状瘤（JLP）的病变组织进行人乳头状瘤病毒（HPV）分型及体液、细胞免疫检测，并对其临床资料进行回顾性分析，以探讨JLP患儿HPV感染类型、途径及细胞、体液免疫水平，提高对其临床特点的认识，探讨有效的治疗方法。     

山东东部57例喉癌人乳头状瘤病毒DNA及其亚型的检测

目的 通过检测山东东部地区喉鳞状细胞癌中人乳头状瘤病毒(HPV)的DNA及其亚型的表达,探讨HPV感染与喉癌的关系及其在喉癌发病中的意义。方法 采用聚合酶链式反应(PCR)扩增和DNA反向点杂交相结合的DNA芯片技术,检测57例喉鳞状细胞瘤各亚型HPV DNA,对照组为同期喉乳头状瘤细胞8例。结果 喉癌组HPV感染率为7.02%(4/57),喉乳头状瘤组75.00%(6/8),差异有统计学意义(χ2=24.906,P＜0.01);喉癌组高危型HPV16与低危型HPV43感染率分别与喉乳头状瘤比较,差异均无统计学意义(χ2= 5.611、0.143,P＞0.05);喉乳头状瘤组低危型HPV11与HPV6感染率较喉癌组高,差异均有统计学意义(χ2=8.611、14.702,P＜0.05)。结论 山东东部地区喉癌患者HPV感染率较低,而低危型HPV6/11与喉乳头状瘤关系较为密切     

HPV6b病毒样颗粒免疫治疗儿童喉乳头状瘤临床初步研究

目的 :研究HPV6bL1病毒样颗粒 (VLP)治疗儿童喉乳头状瘤 (JLP)的安全性和免疫原性。方法 :应用基因工程制备的HPV6bL1VLP 5、10、2 5 μg 3种剂量递增方法对 10例严重复发性JLP患儿进行免疫接种 ,记录不良反应及行血、尿常规和生化检测 ,ELISA法检测血清特异性HPV6bL1VLP抗体 ,对 7例患儿进行迟发性超敏反应 (DTH)试验 ,纤维喉镜随访观察喉部病变情况。结果 :接种后患儿无局部和全身不良反应 ,血清均能产生特异性的中和抗体 ,接种前 3天和 3种剂量完成后及开始治疗 1年后的血清抗体吸收度A均值分别为 0 .110± 0 .0 35 ,0 .310± 0 .0 12 ,0 .5 87± 0 .0 12 ,0 .75 2± 0 .0 19,0 .772± 0 .0 13。第 1剂量完成后与接种前 3天A均值比较 ,第 2剂量与第 1剂量完成后比较 ,第 3剂量与第 2剂量完成后比较 ,接种 1年后与对照组比较 ,各组间差异均有统计学意义 (均P <0 .0 1)。 7例行DTH试验的患儿均呈阳性反应。经免疫治疗后的 10例患儿未见复发。结论 :HPV6bL1VLP对JLP具有安全性和免疫原性 ,可成为防治JLP的有效疫苗。     

显微支撑喉镜下手术摘除加5-FU肿瘤基底部注射治疗喉乳头状瘤36例

目的探讨喉乳头状瘤的治疗方式.方法对1995年1月～2001年12月36例喉乳头状瘤患者在全麻高频通气显微支撑喉镜下手术切除加肿瘤基底部5-氟尿嘧啶(5-FU)局部注射,术后常规给予α-干扰素皮下注射进行回顾分析.结果喉乳头状瘤复发率降低,气管切开后易发生肿瘤种植.结论对喉乳头状瘤的治疗,强调手术摘除加肿瘤基底部5-FU注射是较为有效的方式,治疗中提倡微创,尽量实施内镜下手术,尽量避免喉裂开等手术,以免引起并发症.     

儿童喉乳头状瘤细胞的原代培养及生物学特性研究

目的　探讨人乳头状瘤病毒 (humanpapillomavirus,HPV)感染阳性患儿喉乳头状瘤细胞在体外培养的生物学特性。方法　 2 0 0 0年 3月～ 2 0 0 1年 4月采用组织块培养法培养 10例 12例次喉乳头状瘤患儿手术的标本———HPV感染阳性患儿喉乳头状瘤细胞 ,观察其生长情况 ,计数法绘制细胞生长曲线 ,运用共同引物聚合酶链反应 (polymerasechainreaction ,PCR)法及核酸分子斑点杂交方法对喉乳头状瘤细胞在培养前后的HPVDNA进行检测。结果　HPV感染阳性的喉乳头状瘤细胞在体外生长时间可长达 6周 ,培养前后细胞内均含有HPV的DNA。细胞生长分为 3期 ,延缓期、生长期及停滞期。培养前 1～ 4d细胞从组织块中大量游出 ,5～ 7d为延缓期 ,此期间细胞渐渐开始贴壁 ,8～ 18d为生长期 ,细胞数目迅速增多 ,生长速度很快 ,继而进入停滞期 ,细胞数目增长缓慢 ,细胞空泡化明显 ,渐渐走向死亡。结论　HPV感染阳性的喉乳头状瘤细胞在体外生长情况良好 ,但要建立HPV感染阳性的喉乳头状瘤细胞的动物模型尚需进一步研究     

Effect of human papilloma virus expression on clinical course of laryngeal papilloma

Conclusion. Our observations suggest that human papilloma virus (HPV) 6/11 is the main causative agent of laryngeal papilloma and that detection of active HPV DNA expression may be helpful in identifying patients with aggressive recurrent laryngeal papilloma. Objectives. HPV is assumed to be the main causative agent of this disease. We investigated the expression of the entire genotype of HPV in cases of laryngeal papilloma and correlated their expression with the clinical course of the disease. Subjects and methods. Seventy cases of laryngeal papilloma were evaluated for the presence of the HPV genome by in situ hybridization (ISH) using wide-spectrum HPV DNA probe. Specific types of HPV infection were determined by DNA ISH using type-specific HPV DNA probes (HPV 6, 11, 16, 18, 31, 33). Separate analyses were conducted comparing viral types, frequency of recurrences and duration of disease-free periods. Results. We detected HPV DNA in 40 of the 70 laryngeal papilloma cases (57%). In particular, HPV DNA was detected in 75% of the juvenile types. There were significant associations between HPV and laryngeal papilloma (p<0.01). Among the HPV-positive cases, major specific types were HPV 6/11 (97%). Significant associations were also noted between viral expression and clinical course.     

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。     

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。     

Effect of human papilloma virus expression on clinical course of laryngeal papilloma

Conclusion. Our observations suggest that human papilloma virus (HPV) 6/11 is the main causative agent of laryngeal papilloma and that detection of active HPV DNA expression may be helpful in identifying patients with aggressive recurrent laryngeal papilloma. Objectives. HPV is assumed to be the main causative agent of this disease. We investigated the expression of the entire genotype of HPV in cases of laryngeal papilloma and correlated their expression with the clinical course of the disease. Subjects and methods. Seventy cases of laryngeal papilloma were evaluated for the presence of the HPV genome by in situ hybridization (ISH) using wide-spectrum HPV DNA probe. Specific types of HPV infection were determined by DNA ISH using type-specific HPV DNA probes (HPV 6, 11, 16, 18, 31, 33). Separate analyses were conducted comparing viral types, frequency of recurrences and duration of disease-free periods. Results. We detected HPV DNA in 40 of the 70 laryngeal papilloma cases (57%). In particular, HPV DNA was detected in 75% of the juvenile types. There were significant associations between HPV and laryngeal papilloma (p<0.01). Among the HPV-positive cases, major specific types were HPV 6/11 (97%). Significant associations were also noted between viral expression and clinical course.     

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     

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．     

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、3 3 5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 5 4 2 % (19/ 35 ) ,HPV11型感染率为 2 5 7% (9/ 35 ) ,多重型别HPV6 11感染率为 11 4% (4/ 35 ) ;HPV16、18、3 3 型 ,均为阴性。对照组各型检测结果均为阴性 ,两者对比差异有高度显著性 (P <0 0 0 1)。结论　温州地区婴幼儿咽喉乳头瘤的发生与HPV感染密切相关 ,尤以HPV6感染为主。PCR结合核酸斑点杂交技术检测HPV具有敏感性高、特异性强的优点 ,值得推广。     

Pharyngeal squamous cell papilloma in adult Japanese: comparison with laryngeal papilloma in clinical manifestations and HPV infection

A number of reports have investigated the relationship between laryngeal papilloma and human papilloma virus (HPV) infection. On the other hand, it is unclear whether the HPV infection is involved in the occurrence of pharyngeal papilloma. We hypothesized that HPV infection was involved in the occurrence of pharyngeal papilloma similarly to laryngeal papilloma. To verify this hypothesis, we investigated the presence of HPV infection. Furthermore, clinical manifestations of pharyngeal papilloma, which had rarely been reported, were discussed. A male-to-female ratio, solitary or multiple occurrences, and koilocytosis were examined in cases with pharyngeal papilloma. HPV DNA was examined with unfixed surgically resected specimens of pharyngeal papilloma. A screening test by the liquid-phase hybridization method was carried out for the HPV high-risk group (16, 18, 31, 33, 35, 39, 45, 51, 56, 58, 59, and 68) and HPV low-risk group (6, 11, 42, 43, 44). As a control, 15 cases with laryngeal papilloma for which the same screening test was carried out were employed. Pharyngeal papilloma occurred as a solitary lesion more often, whereas laryngeal papilloma occurred as multiple tumors more frequently. The HPV infection rate was 0% in pharyngeal papilloma cases, which was in stark contrast with 66.7% in the HPV low-risk group in laryngeal papilloma cases. Pharyngeal papilloma occurred as a solitary lesion in females more frequently. Contrary to our hypothesis, the involvement of HPV infection was unlikely in the occurrence of pharyngeal papilloma.     

High incidence of malignant transformation of laryngeal papilloma in Taiwan

OBJECTIVES: Papillomas of the larynx include solitary laryngeal papilloma and recurrent respiratory papillomatosis. This study investigated the incidence of malignant transformation and assessed possible risk factors for laryngeal papillomas. STUDY DESIGN: A prospective, longitudinal study. METHODS: Twenty-six consecutive laryngeal papilloma patients were prospectively studied for 5 or more years, and each patient was periodically examined at 3 to 6 month intervals. A detailed epidemiologic questionnaire was administered at the initial visit. After enrollment, tissue obtained during each laryngeal surgery was examined by polymerase chain reaction assay for human papilloma virus (HPV) and typing. RESULTS: During 237 person-years of follow-up, six new, pathologically confirmed cases of laryngeal carcinoma were ascertained (incidence 2.5/100 person-years), and all were associated with HPV-6 or HPV-11. Malignant transformation revealed no correlation with the following: age less than 3 years at diagnosis, sex, history of tobacco use, history of alcohol consumption, family history of laryngeal cancer, or type of laryngeal papilloma. Laryngeal papilloma without demonstrable HPV DNA was the only significant risk factor for malignant transformation (P < .05). The cumulative risk of malignant transformation in subjects without demonstrable HPV DNA was significantly higher than that in HPV-positive patients (relative risk, 8.0; 95% confidence interval, 1.1-60.3; P = .05). CONCLUSIONS: A relatively high incidence of malignant transformation of laryngeal papilloma was noted in Taiwanese patients. Patients without demonstrable HPV DNA require more frequent follow-up and may benefit from anti-HPV vaccinations.     

Human papilloma viruses in the human hypopharynx

The possible association of human papilloma viruses (HPV) with laryngopharyngeal squamous cell carcinoma is under investigation. Recent work suggests regional differences in the prevalence of HPV infection in the hypopharynx. The present study investigates the prevalence of HPV in tissue obtained from a series of piriform fossae. Piriform fossa epithelium was harvested from 12 autopsy cases free of local disease. DNA was obtained by SDS/Proteinase K digestion. Evidence of HPV infection was documented by the polymerase chain reaction using oligonucleotide primers complementary to sequences in the E6 region of HPV types 11, 16 and 18. All the specimens were positive for beta-globin. HPV11 was isolated from two patients. None were positive for HPV16 or HPV18. An 18% prevalence rate for HPV11 in the normal human hypopharynx was found.