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     

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。     

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。     

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。     

无呼吸道乳头状瘤儿童上呼吸道乳头状瘤病毒的检测

目的 检测儿童扁桃体和腺样体手术标本中的人乳头状瘤病毒(human papillomavirus,HPV)DNA,以了解无复发性呼吸道乳头状瘤(juvenile-onset recurrent respiratory papillomatosis,JORRP)儿童的上呼吸道HPV感染情况.方法 前瞻性研究2008年2月至2009年1月因扁桃体肥大或炎症以及腺样体肥大手术的患儿241例,均确诊为无JORRP或其他已知HPV相关疾病.共取得新鲜的扁桃体标本177个和腺样体标本195个.为验证本研究实验方法的可靠性,取同期17例喉乳头状瘤患儿的新鲜乳头状瘤标本作阳性对照.所有标本采用荧光定量聚合酶链反应法检测HPV DNA并分型.结果 17例乳头状瘤标本HPV-DNA均阳性,均为HPV6或HPV11感染.241例患儿的372个标本中,仅2例扁桃体阳性,分别为HPV6和HPV11感染.再次追问这2例患儿的个人病史和家族史,未发现有HPV感染相关疾病,扁桃体切除样本的病理检查结果均显示淋巴组织增生,无HPV感染的特征病变,也未见恶性肿瘤迹象.本组中上呼吸道HPV感染率为0.8％ (2/241).结论 无JORRP儿童的上呼吸道也存在HPV感染,仅有HPV感染可能不足以引发JORRP.     

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     

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。     

小儿喉乳头状瘤患者的免疫状态调查和局部注射5-氟脲嘧啶的疗效观察

喉乳头状瘤由人乳头状瘤病毒 (HPV)引起 ,以HPV6、1 1型多见。小儿患者发病年龄小 ,病变范围广 ,常侵及喉外组织 ,摘除后易复发 ,而成人患者则易发生癌变。复发性喉乳头状瘤的患儿多伴有体液或细胞免疫功能低下 ,抗病毒能力较差 ,病变范围广且容易侵及下呼吸道 ,使治疗变得尤为困难。 2 0 0 0～ 2 0 0 2年我们收治 9例小儿喉乳头状瘤患者 ,行手术摘除肿瘤后局部注射 5 氟脲嘧啶 ( 5 FU) ,并与 7例不注射 5 FU的对照组进行比较 ,以探讨小儿的免疫状态与肿瘤发生的相关性及局部注射 5 FU的治疗效果。1　资料与方法小儿喉乳头状瘤患者…     

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。     

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     

The detection and significance of human papilloma virus 11b virus like particles and its serum antibody in juvenile larynx papilloma]

OBJECTIVE: To study the relationship between human papilloma virus 11b virus like particles (HPV11bVLPS) serum antibody and the development and prognosis of juvenile larynx papilloma (JLP). METHODS: Enzyme linked immunosorbent assay (ELISA) was used to detect the serum HPV11bVLP antibody (Ab) of 46 JLP's samples in different stage and 20 controls using HPV11bVLPS which was produced by recombinant bacilovirus in insect cells. Grouping: A: control group (n = 20); B: the time of onset was 1 years (n = 15); C: the time of onset was 2 years (n = 15); the patients were followed-up 1 year without recurrence (n = 8); E: The patients were followed-up 2 years without recurrence (n = 8). RESULTS: A value of HPV11bVLP Ab among A, B, C, D, E. group were: (0.073 +/- 0.035); (0.120 +/- 0.049); (0.137 +/- 0.057); (0.518 +/- 0.122); (0.557 +/- 0.144). There was a significant difference between JLP patients and the control group (P < 0.05). The level of HPV11bVLP Ab in (D + E) group (0.534 +/- 0.132) was higher than (B + C) group (0.128 +/- 0.053) (t = 14.90, P < 0.001). CONCLUSION: The results suggested that HPV serum antibody was produced in JLP with HPV infection. There is close relationship between the development and prognosis of the disease and the level of HPV11Ab in serum. The assay of serum HPV11bVLPAb and HPV-VLP could be used as immunological study of HPV11-infection associated disease.     

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。     

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.     

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     

2',5'-Oligoadenylate synthetase activity analysis and human papilloma virus typing as prognostic factors in patients with recurrent respiratory papillomatosis

OBJECTIVE: Determination of early prognostic factors in patients with recurrent respiratory papillomatosis is extremely important, so the major goal of our prospective, multicentre study was to evaluate (1) the feasibility of various factors to determine prognosis of the clinical course, as well as (2) the response to interferon-alpha therapy in recurrent respiratory papillomatosis. METHODS: Forty-two patients with recurrent respiratory papillomatosis were treated with interferon-alpha (3 MU/m(2) three times per week; mean therapy duration was 2.7 +/- 1.8 years) in 1983-1994 and followed-up until 2003. Human papilloma virus (HPV) type, recurrent respiratory papillomatosis severity and 2',5'-oligoadenylate synthetase activity were determined by standard methods and analysed for correlation with the results of long-term clinical outcome. RESULTS AND CONCLUSION: Patients with HPV type 11, a severity score >4, a high number of surgical procedures prior to interferon-alpha therapy and a high basal 2',5'-oligoadenylate synthetase activity should be considered at high risk of an aggressive clinical course, often with spread to lower airway passages, malignant transformation and death. Human papilloma virus type, score for recurrent respiratory papillomatosis severity, number of surgical procedures and 2',5'-oligoadenylate synthetase activity showed significant association with response to interferon-alpha therapy and the long-term clinical course, so these factors have value in predicting prognosis in recurrent respiratory papillomatosis.     

Combined endolaryngeal videoendoscopic surgery and photodynamic treatment of patients with recurrent laryngeal and tracheal papillomatosis

Two-stage combined treatment of chronic recurrent papillomatosis of the larynx and trachea has been designed and tested in P.A. Herzen Research Cancer Institute. Stage I of the treatment consisted in endolaryngeal videoendoscopic surgery with Nd:YAG-laser destruction, argon-plasma coagulation and electroresection of the papilloma; stage II - postoperative photodynamic therapy (PDT) to prevent recurrence. In 1995-2007 the treatment was given to 32 patients aged 10-66 years with recurrent papillomatosis of the airways with the disease history 5-58 years. In two thirds of the patients papillomatosis involved several parts of the larynx and trachea. Squamous cell papilloma was accompanied with dysplasia of the first-second degree in 10 (31%) patients, dysplasia of the third degree - in 4 (12,5%), cancer in situ - in 3 (9,4%) patients. Human papilloma virus (HPV) was detected by hybridization in situ in 96%. The course of the treatment resulted in a complete regression (CR) of papilloma in 25 of 32 (78%), partial regression in 7 patients. The recurrence-free interval averaged 32 months (maximal 7 years) in 14 of 25 patients with CR. HPV was eradicated in a group of patients with persistent clinical remission. A 6 to 19 month follow-up recorded papilloma recurrence in 11 patients. The recurrence-free period increased 2,5-fold. In patients with dysplasia of degree I-III and cancer in situ (n=17) CR of dysplasia and preinvasive cancer foci was achieved in 15 (88%) patients.     

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.