原位免位PCR检测喉癌中人类乳头状瘤病毒E6蛋白

针对常规免疫组化检测石蜡包埋的喉癌组织中人类乳头状瘤病毒（ＨＰＶ）１６／１８型Ｅ６蛋白阳性率低、信号模糊等缺点，我们探索出一种敏感而有效的检测方法———原位免疫ＰＣＲ（ＩｎＳｉｔｕＩｍｍｕｎｏ－ＰＣＲ，ＩＳＩ－ＰＣＲ），报告如下：１　材料与方法１．１　材料：９例１６／１８型ＨＰＶＤＮＡ阳性的喉癌高分化鳞癌石蜡包埋组织。生物素标记ＰａＴＥＬ１４／ＥｃｏＲＩＤＮＡ片断。ＰＣＲ引物：５’－ＡＴＡＣＣＴＡＴＴＧＣＣＴＡＣＧＧＣＡＧ－３’；５’－ＣＧＴＴＡＧＴＡＡＡＴＧＡＡＴＴＴＴＣＴ－３’。１．２　方法…     

嗅神经母细胞瘤的诊断与治疗

目的：探讨嗅神经母细胞瘤的诊断和治疗效果。方法：２２例患者,１５例常规病理检查明确诊断,７例用ＮＳＥ、ＮＦ、ＬＣＡ、ＥＭＡ、ＣＥＡ抗体行免疫组化检查。所有病例分别采用手术、放疗、手术加放疗、手术加放疗和化疗进行治疗。结果：ＮＳＥ阳性率７／７,ＮＦ５／７,其余３种抗体检查均为阴性。１９例手术中１４例发现前颅窝底骨质破坏。随访１７例,３年、５年生存率分别为５９％和３５％。结论：免疫组化检测对本病诊断和鉴别诊断有较大帮助,病变位于鼻腔顶部且前颅窝底骨质破坏提示本病的可能。早期诊断加综合治疗能提高本病生存率。     

分枝杆菌多糖促人LAK细胞增殖及对喉癌HEP－2细胞杀伤活性的研究

为探讨分枝杆菌多糖（ｍｙｃｏｂａｃｔｅｒｉａｌｐｏｌｙｓａｃｃｈａｒｉｄｅｓ，ＭＰＳ）在体外对ＬＡＫ细胞（ｌｙｍ－ｐｈｏｋｉｎｅａｃｔｉｖａｔｅｄｋｉｌｅｒｃｅｌｋｉｎｅ）活性的影响，以提高ＬＡＫ细胞杀瘤活性、减少ＩＬ－２（白细胞介素－２）用量和毒副作用。通过ＭＴＴ法测定不同浓度ＩＬ－２、不同浓度ＭＰＳ对ＬＡＫ细胞促增殖能力的影响；以细胞计数法测定不同浓度ＩＬ－２、ＭＰＳ对ＬＡＫ细胞扩增倍数的影响；应用乳酸脱氢酶法测定不同条件刺激的ＬＡＫ细胞对喉癌ＨＥＰ－２细胞杀伤活性的作用。结果证明小剂量ＩＬ－２（２×１０５Ｕ／Ｌ）诱导ＬＡＫ细胞过程中加入ＭＰＳ０．４ｍｇ／Ｌ能显著增强ＬＡＫ细胞的增殖和杀伤喉癌ＨＥＰ－２细胞的活性。其扩增倍数及杀伤活性均显著超过单纯应用ＩＬ－２（１０６Ｕ／Ｌ）诱导的ＬＡＫ细胞。通过检测不同条件下诱导的ＬＡＫ细胞表型，显示ＭＰＳ联合ＩＬ－２诱导的ＬＡＫ细胞ＣＤ２５百分率明显增高，提示ＭＰＳ可能通过促进ＬＡＫ细胞表面ＩＬ－２膜受体表达，提高ＬＡＫ细胞增殖及杀伤活性。因此，ＭＰＳ可望作为一种新型免疫调节剂用于抗肿瘤治疗     

两种免疫调节剂辅助LAK细胞杀伤喉癌细胞的体外研究

为探讨如何提高白细胞介素－２（ＩＬ－２）／ＬＡＫ细胞过继免疫治疗的抗肿瘤作用，应用免疫调节剂分枝杆菌多糖（Ｍｙｃｏｂａｃｔｅｒｉａｌｐｏｌｙｓａｃｃｈａｒｉｄｅｓ，ＭＰＳ）或ＣＤ３单克隆抗体协同ＩＬ－２诱导的健康人外周血淋巴细胞来源的ＬＡＫ细胞在体外扩增，测定其对喉癌ＨＥＰ－２细胞杀伤活性。实验结果：①不同免疫调节剂对ＬＡＫ细胞扩增的影响：实验组三组细胞于体外扩增均明显高于对照组（Ｐ＜０．０１）；ＭＰＳ组或ＣＤ３组细胞的扩增倍数比单纯应用ＩＬ－２组为高（Ｐ＜０．０１）；其中扩增倍数最高者为ＭＰＳ组，与ＣＤ３组相比，Ｐ＜０．０５。②不同免疫调节剂诱导的ＬＡＫ细胞对喉癌ＨＥＰ－２细胞的杀伤活性比较：在培养一周时，实验组三组对ＨＥＰ－２细胞的杀伤活性均高于对照组，以ＣＤ３组及ＭＰＳ组杀伤活性最高，其杀伤活性均为５５．５％；于培养第二周，ＣＤ３组杀伤ＨＥＰ－２细胞活性仍较高，其他实验组细胞杀伤活性均明显下降。结论：免疫调节剂ＭＰＳ和ＣＤ３单克隆抗体均可通过增强ＬＡＫ细胞体外增殖和杀瘤活性而提高肿瘤过继免疫治疗效果。     

唾液酸与急相蛋白对鼻咽癌的诊断价值

于１９９１年测定了鼻咽癌（ＮＰＣ）和喉癌（ＬＣ）患者血清唾液酸（ＳＡ）和５种急相蛋白（ＡＰＰｓ），其中包括前清蛋白（ＰＡ），ａ＿１－抗胰蛋白酶（ａ＿１－ＡＴ），ａ＿１－酸性糖蛋白（ａ１－ＡＧＰ），触珠蛋白（ＨＰ）和转铁蛋白（ＴＦ），并将上述测定值组合入判别函数式，用于ＮＰＣ和ＬＣ的计算机辅助诊断和随访。本文为验证１９９１年报道的判别函数式，对２３８例未知诊断的初诊患者，双盲法检测血清ＳＡ、５ＡＰＰｓ和鼻咽组织活检及临床检查，结果发现：鼻咽癌的判别函数诊断与临床检查及活检结果符合率均在８０％以上。     

鼻咽泡状核细胞癌和低分化鳞癌的临床生物学行为的对比研究

目的：探讨鼻咽泡状核细胞癌（ＶＮＣＣ）和低分化鳞癌（ＰＤＳＣＣ）的临床生物学行为的差异性。方法：用原位杂交和免疫组化染色（ＳＰ法）技术,检测ＶＮＣＣ和ＰＤＳＣＣ癌组织中ＣｅｒｂＢ２、表皮生长因子受体（ＥＧＦＲ）和增殖细胞核抗原（ＰＣＮＡ）表达,用原位末端标记（ＩＳＬＥ）方法检测癌细胞凋亡,并对其预后和临床特征进行对比分析。结果：①ＶＮＣＣ的ＩＳＥＬ和ＰＣＮＡ染色强度指数（ＳＩ）明显高于ＰＤＳＣＣ;②ＶＮＣＣ的ＣｅｒｂＢ２、ＥＧＦＲｍＲＮＡ和ＣｅｒｂＢ２蛋白表达与ＰＤＳＣＣ无明显差别;③ＶＮＣＣ的５年生存率比ＰＤＳＣＣ高。结论：与ＰＤＳＣＣ比较,ＶＮＣＣ是一种生长增殖活跃、分化程度更低,但有较好预后趋向的一种癌型。     

正常清醒豚鼠的畸变产物耳声发射特性

目的 研究正常清醒豚鼠的畸变产物耳声发射（ＤＰＯＡＥ）的特性。方法 采用ＣＥＬＥＳＴＡ ５０３型耳声发射分析仪对２６只正常清醒豚鼠（３５耳）进行ＤＰ图及ＤＰ输入／输出曲线（ＤＰ－Ｉ／Ｏ）的测试，随机选择１１只正常清醒豚鼠（２０耳）进行ＤＰＯＡＥ的重复测试，用ＳＰＳＳ１０．０对数据进行统计分析。结果 在ＤＰ图中，当初始音强度Ｌ１／Ｌ２为 ７０／６５ ｄＢ ＳＰＬ时，正常清醒豚鼠的 ＤＰＯＡＥ幅值随测试频率ｆ０从０．７５－８ｋＨｚ的增加而逐渐升高（２７．９０±１．９６－5０．６５±０．７１）。在 ＤＰ－Ｉ／Ｏ中，当ｆ０分别为４、６、８ ｋＨｚ时，正常清醒豚鼠的ＤＰＯＡＥ幅值随Ｌ１／Ｌ２从７０／６５以５ｄＢ－挡降至１５／１０ ｄＢ ＳＰＬ而呈线性下降（Ｐ＜０．０１），在Ｌ１／Ｌ２为５５／５０或６０／５５ ｄＢ ＳＰＬ处出现饱和或低谷，同一Ｉ／Ｏ曲线上Ｌ１／Ｌ２分别从７０／６５及５５／５０ ｄＢ ＳＰＬ递减至阈值的Ｉ／Ｏ斜率（分别记为ＫＴ及ＫＬ）均接近于１，且ＫＬ大于ＫＴ（Ｐ＜０．０１）。重复测试的ＤＰＯＡＥ幅值差异小（＜ １ｄＢ ＳＰＬ）且无统计学意义（Ｐ＞０．０５）。结论 正常清醒豚鼠ＤＰＯＡＥ测试充分表现了其捡出率高、反应幅值大     

鼻T／NK细胞淋巴瘤14例探讨

目的 探讨鼻Ｔ／ＮＫ细胞淋巴瘤的临床特点、病理诊断标准和生物学行为，为诊断和治疗提供经验。方法 选择既往诊断为“恶性肉芽肿”及相关疾病２３例：①复习病理切片、进行免疫组化及ＥＢ病毒原位杂交染色，选用的抗体有：ＴＩＡ－１、ＣＤ４３、ＣＤ２０、ＣＤ３ε、ＣＤ５６，原位杂交选用ＦＴＴＣ标记，针对ＥＢ病毒编码的小分子ＲＮＡ的寡核酸探针（ＥＢ ｖｉｒｕｓ ｅｎｃｏｄｅｄ ｓｍａｌｌ ｎｕｃｌｅａｒ ＲＮＡ，ＥＢＥＲ原位杂交）；②复习病历，总结临床病理特点。结果 ①２３例中１４例符合鼻Ｔ／ＮＫ细胞淋巴瘤形态学改变，免疫组化显示肿瘤细胞不表达ＣＤ２０，表达ＴＩＡ－１、ＣＤ５６、ＣＤ３ε或ＣＤ４３，ＥＢＥＲ原位杂交阳性；②１４例患者中８例侵犯鼻部中线，其余分别原发于鼻咽、咽、喉。其中２例累及消化道，１例合并嗜血综合征。结论 ①     

γ—干扰素对喉癌细胞株HEP—2增殖活性的影响

用单克隆抗体Ｋｉ－６７（抗ＰＣＮＡ），以链霉菌素－生物素技术（ＬＳＡＢ）检测喉癌细胞株ＨＥＰ－２增殖细胞核抗原（ＰＣＮＡ）表达。人重组γ－干扰素（ｒｈｕ－ＩＦＮ－γ）抑制ＰＣＮＡ表达（即抗增殖活性）强弱与其剂量有关；雌激素对ｒｈｕ－ＩＦＮ－γ抗增殖作用无影响。提示：ｒｈｕ－ＩＦＮ－γ对喉癌细胞株ＨＥＰ－２有抗增殖作用，因而在喉癌的治疗中具有应用价值的。     

增殖细胞核抗原在喉上皮癌前病变中的表达及意义

用免疫组化技术对１１例喉上皮单纯性增生（ＳＨＥ），３２例非典型性增生（ＡＨＥ）及４２例喉鳞状细胞癌（ＬＳＣＣ）的增殖细胞核抗原（ＰＣＮＡ）表达进行了原位检测。结果表明；在ＳＨＥ、ＡＨＥ及ＬＳＣＣ中，ＰＣＮＡ指数分别为９．５７％、２７．３３％、６８．０５％，差异极有显著性意义（Ｐ＜０．００１）。在轻、中、重度ＡＨＥ中，ＰＣＮＡ指数分别为１３．７９％、３０．８４％、３９．９４％，差异有非常显著性意义（Ｐ＜０．０１）。提示ＰＣＮＡ表达程度与非典型性增生程度密切相关，是反映细胞增殖状态的生物标志，ＰＣＮＡ组化研究有助于认识喉癌前病变的本质，发展趋势及喉癌的早期诊断。     

The expression of vascular endothelial growth factor (VEGF) and VEGF‐C in early laryngeal cancer: relationship with radioresistance

The expression of vascular endothelial growth factor (VEGF) and VEGF‐C in early laryngeal cancer: relationship with radioresistance Angiogenesis is essential for tumour growth and invasion. Vascular endothelial growth factor (VEGF) is a prime mediator of tumour angiogenesis. VEGF‐C is a closely related protein that effects lymphatic endothelial cells and may be important in the process of lymphatic metastasis. The purpose of this study was to evaluate the expression of these cytokines in patients with T1 and T2a glottic, squamous cell carcinoma, in comparison with normal epithelial control tissue, to ascertain any association with radioresistance. Twenty‐two tumours treated by radiotherapy (13 radiosensitive, nine radioresistant) and seven normal control tissues were studied. The minimum follow‐up was 2 years after radiotherapy. Expression of VEGF and VEGF‐C was evaluated by immunohistochemistry of formalin‐fixed, paraffin‐embedded biopsy specimens. Analysis was carried out using a quantitative computer image analyser. Both VEGF and VEGF‐C were detectable in tumour and normal control specimens. There was increased expression in tumour specimens of both VEGF (P = 0.03) and VEGF‐C (P < 0.001). In addition, the expression of VEGF‐C was associated with tumours of higher histological grade (P = 0.021). There was, however, no difference in VEGF and VEGF‐C expression between radioresistant and radiosensitive tumours. The expression of VEGF and VEGF‐C is increased in early laryngeal squamous cell carcinoma (SCC). However, measuring the expression of these proteins cannot predict radioresistance in this tumour group.     

Treatment of Maxillary Sinus Carcinoma: Clinical Results Using the Kitasato Modality

The conventional therapeutic regimen for maxillary sinus carcinoma consists of dissection of the maxilla, full-dose irradiation and extensive chemotherapy. However, the results obtained with this treatment are often poor. Even when patients recover, their quality of life is significantly reduced as a result of deformity of facial structures and swallowing and articulation dysfunctions. A retrospective analysis of 68 patients with maxillary sinus carcinoma treated with the Kitasato modality between 1975 and 1999 was conducted. All patients underwent pergingival maxillary sinus surgery combined with pre- and postoperative irradiation therapy with standardized total doses of 16 Gy; the postoperative irradiation was given in combination with regional intra-arterial infusion chemotherapy administered via the superficial temporal artery. All visible tumor lesions were removed where possible in order to preserve or facilitate cellular immunity after surgery. The cumulative 5-year survival rates were 85.7% for Stage II patients, 88.1% for Stage III, 76.6% for Stage IVA and 75.0% for Stage IVB.     

Tongue Base Reduction with Temperature-controlled Radiofrequency Volumetric Tissue Reduction for Treatment of Obstructive Sleep Apnea Syndrome

In recent years a considerable effort has been made to establish the use of different surgical techniques for the treatment of obstructive sleep apnea syndrome (OSAS). Nevertheless, treatment of hypopharyngeal obstruction due to tongue base hypertrophy remains in many ways an unsolved problem. The aim of this study was to evaluate the safety and efficacy of tongue base reduction with temperature-controlled radiofrequency volumetric tissue reduction in the treatment of OSAS. Twenty patients with OSAS and tongue base hypertrophy were treated with radiofrequency tissue ablation. An intensified treatment protocol was used, delivering 2,800 J per treatment session under local anesthesia. Two nights of polysomnography testing were performed before and after treatment. Daytime sleepiness, snoring and postoperative morbidity were assessed using questionnaires. Mean respiratory disturbance index (RDI) was reduced from 32.1 to 24.9/h after a mean of 3.4 treatment sessions. Six patients (33%) were cured after the procedure (reduction in RDI of &#83 50% and a postoperative RDI of <15/h) and ten (55%) showed an improvement of >20% in their RDI. Daytime sleepiness and snoring improved significantly. Peri- and postoperative morbidity was low; one severe complication occurred (tongue base abscess). We were able to achieve similar cure and responder rates to those reported in a recently published pilot study but with a reduced number of treatment sessions. We believe that this technique may improve patient acceptance and have beneficial cost implications.     

Cervical Reflex Induced by Click Stimuli in Cats

We studied click-evoked potentials in the anterior horn of the spinal cord in 17 cats. A concentric needle electrode was inserted into the anterior horn of the spinal cord at levels C3-C6. Potentials evoked with 105 dB SPL clicks were recorded with a peak latency of 4.89-5.10 ms only at the C3 level. These responses were observed 45-60 dB SPL above the auditory brainstem response (ABR) threshold, and no potentials were evoked by stimulation of the contralateral ear. Average was performed 100 times with changes in stimulation frequency of 1-20 Hz. The amplitude of the potentials decreased with increasing stimulus frequency, but there were no changes in ABRs. The responses disappeared after destruction of the medial vestibulospinal tract at the obex level, but ABRs were still recorded. The spinal nucleus of the accessory nerves was located in the anterior horn of the spinal cord at levels C1-C6, and the sternocleidomastoid muscle motoneurons were found at levels C1-C3. The click-evoked potentials recorded in this study reflect responses of the spinal nucleus of accessory nerves through the vestibulospinal tract to click stimulation. The responses have the same characteristics as vestibular-evoked myogenic potentials that can be recorded using surface electrodes over the sternocleidomastoid muscles of humans.     

Lack of significant estrogen and progesterone receptor expression in nasal telangiectasias in hereditary hemorrhagic telangiectasia: an immunohistochemical analysis

Obstructive sleep apnea syndrome (OSAS) is characterized by snoring and apnea during sleep leading to decreased oxygen saturation and disturbed sleep, excessive daytime sleepiness and neuropsychological disturbances. This study investigates cognitive neuropsychological abilities in a group of 53 OSAS patients before and after treatment with uvulopalatopharyngoplasty. General intellectual ability, verbal learning and memory as well as executive functioning were measured at baseline and 6 months postoperatively. After surgery there were significant improvements in verbal learning and memory (mean change - 39, SD 57.3, p <0.001), recall (mean change - 24.3, SD 39.3, p <0.001) and executive functioning (as assessed by percentage of errors on the Wisconsin Card Sorting Test; mean change-9.1, SD 15.7, p <0.001). These improvements were in accordance with improvements in the degree of sleep apnea, the oxygen desaturation index (mean change -9.7, SD 15.9, p <0.001) and arterial minimum oxygen saturation (mean change 4.5%, SD 10.2%, p <0.01). Surgical treatment seems to improve verbal learning, memory and recall and executive functions in parallel with better oxygenation in OSAS.     

Measurement of chemosensory function

Although hundreds of thousands of patients seek medical help annually for disorders of taste and smell, relatively few medical practitioners quantitatively test their patients' chemosensory function, taking their complaints at face value. This is clearly not the approach paid to patients complaining of visual, hearing, or balance problems. Accurate chemosensory testing is essential to establish the nature, degree, and veracity of a patient's complaint, as well as to aid in counseling and in monitoring the effectiveness of treatment strategies and decisions. In many cases, patients perseverate on chemosensory loss that objective assessment demonstrates has resolved. In other cases, patients are malingering. Olfactory testing is critical for not only establishing the validity and degree of the chemosensory dysfunction, but for helping patients place their dysfunction into perspective relative to the function of their peer group. It is well established, for example, that olfactory dysfunction is the rule, rather than the exception, in members of the older population. Moreover, it is now apparent that such dysfunction can be an early sign of neurodegenerative diseases such as Alzheimer's and Parkinson's. Importantly, older anosmics are three times more likely to die over the course of an ensuring five-year period than their normosmic peers, a situation that may be averted in some cases by appropriate nutritional and safety counseling. This review provides the clinician, as well as the academic and industrial researcher, with an overview of the available means for accurately assessing smell and taste function, including up-to-date information and normative data for advances in this field.