鼻咽癌FRA3B脆性部位的杂合性丢失与临床病理特征的关系

目的 研究鼻咽癌染色体脆性部位FRA3B区域的等位基因杂合性丢失（loss of heterozygosity，LOH）情况，并探讨LOH与鼻咽癌临床病理特征及EB病毒（Epstein-Barr virus，EBV）感染的关系。方法 选择FRA3B区域的8个微卫星多态标记对40例鼻咽癌组织进行LOH分析。结果 77．5％（31／40）的鼻咽癌在FRA3B脆性部位出现LOH，丢失频率较高的2个位点是D3S1300（55．6％）和D3S2757（50．0％），其共同缺失区为D3S4103-D3S4260。LOH与鼻咽癌的临床病理特征（肿瘤T分期、颈淋巴结转移、临床分期、肿瘤分化程度、肿瘤复发情况及EBV壳抗原IgA抗体滴度）相关，临床Ⅲ～Ⅳ期、低分化鳞癌、肿瘤复发和抗体滴度≥1：40者LOH频率明显高于临床Ⅰ～Ⅱ期、中分化鳞癌、无肿瘤复发及抗体滴度〈1：40者（P〈0．05）。结论 FRA3B脆性部位的LOH为鼻咽癌的频发事件，可能参与了鼻咽癌的发生发展，共同缺失区D3S4103-D3S4260可能是其优先作用的分子靶点。     

鼻咽癌染色体3p14的精细等位基因缺失分析

目的：进一步明确鼻咽癌染色体3p14区区域等位基因杂合性丢失（loss of heterozygosity,LOH)的频率与共同缺失区规范,以便分离该区域内与鼻咽癌相关的候选抑瘤基因。方法：选择位于3p14的6个高密度微卫星多态标记,对32例鼻咽癌组织进行LOH分析。结果：71．88％（23／31）的鼻咽癌在至少1个位点发生LOH,丢失频率较高的3个位点是D3S1313（46．43％）、D3S1300（50．0％）和D3S1312（44．44％）,在存在丢失的23例患者中,8例表现为一个连续的非随机的LOH区域,其最小共同缺失区为D3S1313－D3S1312（约3．4个厘摩）。且该区域的缺失与鼻咽癌临床分期、EB病毒感染有明显关系。结论：鼻咽癌在3p14的最小共同缺失区位于D3S1313－D3S1312之间,该区域可能存在一个尚克隆的与鼻咽癌发生发展密切相关的抑瘤基因。     

微卫星不稳定和错配修复基因异常在喉鳞状细胞癌中相关性研究

目的 研究喉鳞状细胞癌中微卫星不稳定（microsatellite instability,MSI）发生的临床意义及其与错配修复基因（mismatch repair gene,MMR）表达的相关性。方法 50例喉鳞状细胞癌患者的石蜡切片选自北京同仁医院2002年至2003年的手术标本,利用显微切割-多聚酶链反应-单链长度多态性分析-银染的方法进行MSI的检测,统计MSI的发生率及其与临床资料的相关性,应用免疫组织化学观察MMR中hMLH1和hMSH2的表达。五个微卫星位点位于染色体1P,3p,5q,9p,17p上,分别临近BCAR3（breast cancer anti-estrogen resistance3）,FHIT,APC,CDKN2A（p16）,TP53等基因。结果 在五个微卫星位点（D17S796,D3S3544,D5S656,D1S375,D9S162）提供统计信息的病例数分别是44,42,45,44和40例。MSI的发生率低于杂合性缺失（loss of heterozygosity,LOH）的发生率。MSI的发生率分别是：D17S796（TP53）20．5％（9／44）,D3S3544（FHIT）14．3％（6／42）,D5S65631．1％（14／45）,D1S375（BCAR3）20．5％（9／44）,D9S162（CDKN2A）15．0％（6／401。MSI的发生与年龄、性别、吸烟史、肿瘤部位、肿瘤分化、TNM分期的关系没有统计学意义（P〉0．05）,但是与肿瘤复发的相关性具有统计学意义（P〈0．01）。MSI的发生与MMR的表达存在相关性（P〈0．01）。MMR阳性细胞和阴性细胞共存在同一张切片内是MMR免疫组织化学的特点。结论 微卫星不稳定和错配修复基因异常可能参与部分喉鳞状细胞癌的发生,微卫星不稳定可能是喉鳞癌复发的特征性指标。     

喉癌及癌前病变等位基因不平衡性的相关研究

目的探讨喉鳞状细胞癌变过程中微卫星DNA等位基因不平衡性的特征及其意义。方法选取染色体3P、9P和17P上6个多态性微卫星位点D3S1234、D9S171、D9S1748、D9S162、INFA和D17S796,利用聚合酶链式反应一简单序列长度多态性一银染技术,对49例喉癌癌前病变和喉癌组织进行等位基因不平衡分析,统计杂合性缺失（10ssofheterozygosity,LOH）和微卫星不稳定性（microsatelliteinstability,MSI）的发生率及其与临床病理特征的相关性。结果6个微卫星标记物LOH和MSI发生率分别为：喉癌癌前病变中单纯过度增生为3．7％和14．8％,轻度不典型增生为10．8％和21．6％,重度不典型增生为26．0％和23．3％;喉鳞状细胞癌为38．7％和21．3％。其中LOH的总检出率在不同病理组间有统计学意义（X2=17．686,P=0．000）,而MSI的检出率统计学意义（X2=0．314,P〉0．05）。不同病理组间D9S171和D9S162单个位点LOH检出率有统计学意义（P=0．022,P=0．025）。在癌前病变早期MSI发生率高于LOH。结论等位基因不平衡可能参与喉癌发生发展,微卫星分析法为喉癌癌前病变的早期诊断提供新的途径。     

喉鳞状细胞癌中微卫星的异常改变

目的：从分子生物学水平探究微卫星的不稳定性（MSI）与杂合性缺失（LOH）在喉鳞状细胞癌发病机制中的意义。方法：选择3号,5号及11号染色体的3个微卫星位点采用PCR和聚丙烯酰胺凝胶电泳-硝酸银染色方法对40例喉鳞状细胞癌患者手术切除的癌组织及癌旁正常组织进行微卫星分析。结果：40例喉鳞状细胞癌中,35例（87．5％）分别有1～3个微卫星位点发生MSI或LOH。微卫星异常改变发生率最高的位点为D5s592,占70％（28／40）;其次是D3s1228位点,占52．5％（21／40）。结论：在3p14区域及5q23区域附近的抑癌基因参与致癌机制,D3s1228和D5s592的微卫星改变与喉鳞状细胞癌的临床分期相关。     

喉癌、喉咽癌脆性组氨酸三联体基因杂合性缺失和微卫星不稳定

目的 探讨脆性组氨酸三联体(fragile histidine triad,FHIT)基因杂合性缺失(loss of heterozygosity,LOH)和微卫星不稳定(microsatellite instability,MSI)与喉癌、喉咽癌发生发展的关系.方法 选取FHIT基因内的三个微卫星位点D3S1234、D3S4103、D3S1300进行PCR扩增,应用单链长度多态性分析-银染技术进行LOH和MSI分析.结果 ①D3S4103、D3S1234、D3S1 300的杂合率分别为76.32%、71.05%和78.95%;②喉癌、喉咽癌总LOH发生率分别是:D3S4103(48.28%)、D3S1234(37.04%)、D3S1300(33.33%);③喉癌、喉咽癌D3S1234、D3S1300、D3S4103的总LOH发生率与患者年龄、性别、吸烟、肿瘤分化程度、肿瘤部位、T分期和肿瘤复发与否的关系均无统计学意义(P＞0.05);④喉癌、喉咽癌总MSI发生率分别是:D3S1234(18.42%)、D3S1300(28.95%)、D3S4103(21.05%);⑤喉癌、喉咽癌FHIT基因D3S1234、D3S1 300、D3S41 03的总MSI发生率与患者年龄、性别、吸烟史、肿瘤部位、肿瘤分化程度和T分期的关系无统计学意义(P＞0.05);⑥复发喉癌、喉咽癌病例FHIT基因总MSI发生率为83.33%,原发病例总MSI发生率为30.77%,复发病例MSI阳性率显著高于原发病例,差异具有统计学意义(P=0.004).结论 ①FHIT基因在喉癌、喉咽癌中既存在杂合性丢失,又存在微卫星不稳定,但以前者为主;②FHIT基因参与了喉癌、喉咽癌的发生,可能是喉癌、喉咽癌候选抑癌基因之一;③FHIT基因微卫星不稳定与喉癌、喉咽癌复发可能相关.     

微切割喉鳞状细胞癌9p13—23区域微卫星杂合性缺失的研究

目的：探讨喉鳞状细胞癌（简称鳞癌）在9p13-23区域微卫星（microsatellite)发生杂合性缺失（loss of heterozygosity,LOH)的热点。方法：采用显微切割法从病理切片中挑取肿瘤组织,选取位于9p13-23区域的13个高多态性微卫星引物对42例喉鳞癌组织进行聚合酶链反应和变性凝胶电泳。结果：（1）42例喉鳞癌在9p13-23区域等位基因LOH的总发生率是97．6％（41／42）。在13个微卫星引物中,LOH发生率最高者是位于9p22-23的D9S162（89．5％）,其次是位于9p21的D9S171（80．0％）,与p16基因紧密连锁的D9S1748的LOH发生率仅50．0％,（2）等位基因缺失作图分析发现42例喉鳞癌组织在9p13-23上存在2个明显的LOH较小区域,分别位于9p21的D9S161－D9S171之间和9p22-23的IFNA和D9S162之间。结论：喉鳞癌在9p13-23区域除抑癌基因p16以外可能还存在2个或2个以上候选抑癌基因,这些候选抑癌基因也许和p16-一样与喉鳞癌的发生,发展密切相关。     

鼻咽癌染色体3p14的精细等位基因缺失研究

目的　研究鼻咽癌染色体 3p14区域的精细等位基因杂合性丢失 (lossofheterozygosity,LOH)情况 ,并探讨LOH与鼻咽癌临床分期、临床病理和EB病毒 (Epstein Barrvirus,EBV)感染的关系。方法　采用 3p14区域 6个精确高密度的微卫星多态性位点 ,对 32例患者鼻咽癌组织进行LOH分析。结果　 32例患者中有 2 3例 ( 71 9% )在至少 1个位点发生LOH ,丢失频率较高的 3个位点是D3S1313( 46 4% )、D3S130 0 ( 5 0 0 % )和D3S1312 ( 44 4% )。在具有丢失的 2 3例患者中 ,12例表现为一个连续的非随机的LOH区域 ,其最小共同缺失区为D3S1313～D3S1312。该区域的LOH与临床分期、EBV感染有明显关系。 30例低分化鳞癌的LOH频率为 70 0 % ,2例泡状核细胞癌均存在 2个位点的LOH。结论　鼻咽癌染色体 3p14区存在较高的LOH率 ,提示在D3S1313和D3S1312之间可能存在尚未克隆的与鼻咽癌发生发展相关的抑癌基因。     

鼻咽癌中VEGF、bFGF的表达及其与颈淋巴结转移的关系

目的 探讨鼻咽癌中血管生长因子VEGF、bFGF的表达及其与颈淋巴结转移的关系。方法 采用免疫组化S－P法对35例鼻咽癌（其中15例同时取颈部转移癌组织）,20例鼻咽粘膜不典型增生组织和10例慢性炎症组织中VEGF、bFGF的表达进行了检测。结果（1）35例鼻咽癌中17例（48．6％）VEGF表达阳性,明显高于非癌对照组（6．7％,P＝0．000）,有颈淋巴结转移鼻咽癌中VEGF的表达（73．3％）明显高于无颈淋巴结转移者（30％,P＝0．018）;晚期（Ⅲ、Ⅳ期,70．6％）明显高于早期（Ⅰ、Ⅱ期,27．8％,P＝0．018）：但咽癌原发灶中VEGF的表达与颈部转移灶中的表达无相关性（P＞0．05）;（2）35例鼻咽癌中有8例（22．9％）bFGF表达阳性,明显高于非癌对照组（3．3％,P＝0．031）。但鼻咽癌中bFGF的表达与患者的年龄、性别、肿瘤的组织学类型和分期无关,也与肿瘤的N分期无关（P＞0．05）。结论 （1）VEGF与鼻咽癌的生长及颈淋巴结转移有关,它可能可以作为预测鼻咽癌颈淋巴结转移的的指标;（2）bFGF可能与鼻咽癌的生长有关系,但与肿瘤的颈淋巴结转移可能无关。     

鼻咽癌染色体3p14的精细等位基因缺失研究

目的 研究鼻咽癌染色体３ｐ１４区域的精细等位基因杂合性丢失（ｌｏｓｓ ｏｆ ｈｅｔｅｒｏｚｙｇｏｓｉｔｙ,ＬＯＨ）情况,并探讨ＬＯＨ与鼻咽癌临床分期、临床病理和ＥＢ病毒（Ｅｐｓｔｅｉｎ－Ｂａｒｒ ｖｉｒｕｓ,ＥＢＶ）感染的关系。方法 采用３ｐ１４区域６个精确高密度的微卫星多态性位点,对３２例患者鼻咽癌组织进行ＬＯＨ分析。结果 ３２例患者中有２３例（７１．９％）在至少１个位点发生ＬＯＨ,丢失频率较高     

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.