喉癌人类组织相容性抗原Ⅰ缺失与癌组织中淋巴细胞浸润的研究

目的检测喉癌组织人类主要组织相容性抗原(human leucotyte antigen,HLA)表达缺失情况,探讨其表达与肿瘤浸润淋巴细胞(tumor infiltrating lymphocyte,TIL)及肿瘤细胞恶性度的关系. 方法 2001年2月～2001年5月应用特异性鼠抗人HLAⅠ类分子等单克隆抗体与喉癌组织结合,通过免疫组化法,对1993～1998年64例手术切除喉癌标本进行检测.结果 64例喉癌组织中有57例HLAⅠ抗原阳性表达,7例阴性表达,阴性表达率为10.9%(7/64);HLAⅠ抗原阳性组织,浸润到肿瘤实质的CD+3T细胞和CD+8T细胞数量明显多于HLAⅠ抗原阴性组织;随着HLAⅠ抗原表达下调,肿瘤细胞恶性度增加.随访52例,3年生存率与HLAⅠ抗原缺失无关(P>0.05).结论部分喉癌组织存在HLAⅠ抗原丢失,且其丢失不利于细胞毒性T淋巴细胞向肿瘤实质浸润与杀伤.HLAⅠ抗原表达缺如,可能促进了细胞恶变的发生发展,是肿瘤逃避免疫监视的方式之一.     

人类白细胞抗原与喉癌的研究

随着分子生物学的发展，已经证实恶性肿瘤的发生、发展和转移是其逃脱机体免疫监控的结果，机体免疫系统主要通过识别肿瘤细胞表面的人类白细胞抗原（Human Leukocyte Antigen，HLA）而发挥其监视作用。喉癌是头颈肿瘤的高发病，目前对于人类白细胞抗原的多态性和人类白细胞抗原异常表达与喉癌的相关性的研究成为热点之一。     

喉癌患者肿瘤浸润淋巴细胞抗肿瘤活性的研究

采用双梯度不连续密度梯度离心的方法，由喉鳞状细胞癌组织中分离出肿瘤浸润性淋巴细胞（ｔｕｍｏｒ ｉｎｆｉｌｔｒａｔｉｎｇ ｌｙｍｐｈｏｃｙｔｅ，ＴＩＬ），经重组人类白细胞介素－２（ｒｅｔｕｒｎ ｉｎｔｅｒｌｅｕｋｉｎ－２，ｒＩＬ－２）激活，成为活化的ＴＩＬ细胞。采用＾５１Ｃｒ体外释放的方法，检查活化的ＴＩＬ细胞对自体肿瘤细胞、异体肿瘤细胞的杀伤特性；并观察了ＴＩＬ细胞在裸鼠体内对自体肿瘤细胞生长的抑     

喉鳞癌组织中浸润淋巴细胞超微结构研究

对１４例喉鳞状细胞癌组织中淋巴细胞（ＴＩＬ）浸润状况及其与癌细胞间的相互关系进行透射电镜观察。结果显示喉鳞癌组织中普通存在着程度不等的淋巴细胞浸润，但多数表现为静止状态，少部分具有活化的母细胞特征且与癌细胞密切接触，使其自身和癌细胞均表现出明显的形态学变化，表明其可探讨ＴＩＬ对肿瘤细胞生长及杀伤机理提供重要的形态学依据。     

基质金属蛋白酶与喉癌、下咽癌的浸润转移

侵袭转移是喉癌、下咽癌的主要生物学行为之一，也是喉癌、下咽癌患致死的主要原因。研究喉癌、下咽癌的侵袭和转移对喉癌、下咽癌的治疗有着重要的意义。基质金属蛋白酶(matriKmettallo—proteinases，MMPs)在肿瘤侵袭和转移中发挥着重要作用，不同的MMPs在喉癌、下咽癌分布和表达不同，因而对喉癌、下咽癌的侵袭和转移的作用也不同。不同的MMPs在喉癌、下咽癌分布和表达不同，有可能成为探索个体化治疗喉癌、下咽癌的理论依据。     

p27和CyclinD1及增殖细胞核抗原在喉癌组织中的表达及其意义

应用免疫组化链霉素抗生物素蛋白-生物素免疫染色法(strept avidin—biotin immunostaining，S-Pmethod，SP)研究p27(cyclin—dependent kinase inhibitor)、CyclinD1、增殖细胞核抗原(proliferating cell nuclear antigen，PCNA)在喉癌中的表达及意义。     

喉癌及癌旁组织中脂质过氧化物及超氧化物歧化酶的研究

本文应用黄嘌呤氧化酶法和硫代巴比妥酸法对３２例喉癌及癌旁组织，１２例正常喉组织中自由基清除酶———超氧化物歧化酶（ＳＯＤ）活性和自由基代谢产物过氧化脂质（ＬＰＯ）含量测定。检测结果表明，喉癌组织中ＳＯＤ活性及ＬＰＯ含量均高于癌旁组织及正常对照组，统计学处理Ｐ＜０．０１，差异非常显著，故认为喉癌的发生与自由基的关系较为密切。     

肿瘤浸润T淋巴细胞对喉鳞状细胞癌预后评估作用的研究进展

喉鳞状细胞癌（LSCC）是常见的头颈部恶性肿瘤之一,近年来发病率有逐渐增长的趋势,但患者5年生存率并无上升趋势,亟需更加稳定有效的预后评估指标来指导治疗、判断预后。肿瘤浸润淋巴细胞（TILs）是一种存在于肿瘤组织内部具有高度异质性的淋巴细胞,以T淋巴细胞为主,在宿主抗原特异性肿瘤免疫应答中发挥关键作用。多项研究表明,TILs与LSCC患者预后密切相关。本文对TILs的一般特征、评估方法及肿瘤浸润T淋巴细胞对LSCC预后判断价值的研究进展进行综述。     

梅尼埃病患者与组织相容性白细胞抗原—Ⅱ类基因关联的研究

OBJECTIVE: To investigate the relation between histocompatibility leukocyte antigen (HLA)-class II DRB1 and Meniére's disease in Chinese. METHODS: Polymerase chain reaction-sequence specific primers (PCR-SSP) technique was used. HLA-class II allele distribution were measured in 60 patients with Meniére's disease, in comparison with those in 85 normal population. RESULTS: The results showed that the frequency of DRB1*09 allele was significantly lower in Meniére's disease patients than in those of controls (relative risk = 0.17, P < 0.01). CONCLUSION: HLA-DRB1*09 of the patients with Meniére's disease was significant low. It implied that DRB1*09 may be a protective gene for Meniére's disease.     

人喉癌引流淋巴结淋巴细胞局部应用疗效观察

目的 为提高肿瘤引流淋巴结淋巴细胞（ＴＤＬＮＬ）转输后在肿瘤局部的数量,采用肿瘤局部注射的方法对喉癌裸鼠移植瘤模型进行实验性治疗。方法 用２０只人喉癌裸鼠移植瘤系ＰＨＣ３的裸鼠移植模型作为治疗对象,随机分为２组。１０只作为治疗组,取新鲜喉癌患者切除的淋巴结,分离得到淋巴细胞,体外培养并用白介素２（ＩＬ－２）１０００ＩＵ／ｍｌ激活后,每周一次注射于瘤内及周围,每次每只注射１０＾７个细胞（悬于０．１ｍ     

人喉癌引流淋巴结淋巴细胞局部应用疗效观察

目的　为提高肿瘤引流淋巴结淋巴细胞（ｔｕｍｏｒ ｄｒａｉｎｉｎｇｌｙｍｐｈ ｎｏｄｅｌｙｍｐｈｏｃｙｔｅ,ＴＤＬＮＬ） 转输后在肿瘤局部的数量,采用肿瘤局部注射的方法对喉癌裸鼠移植瘤模型进行实验性治疗。方法　用２０ 只人喉癌裸鼠移植瘤系ＰＨＣ３ 的裸鼠移植模型作为治疗对象,随机分为２ 组。１０ 只作为治疗组,取新鲜喉癌患者切除的淋巴结,分离得到淋巴细胞,体外培养并用白介素２（ｉｎｔｅｒｌｅｕｋｉｎ２ ,ＩＬ２）１ ０００ＩＵ／ｍｌ 激活后,每周一次注射于瘤内及周围,每次每只注射１０７ 个细胞（悬于０．１ ｍｌ ＤＨａｎｋｓ 液中）,另１０ 只作为对照组,每周一次注射０．１ ｍｌ ＤＨａｎｋｓ 缓冲液于瘤内及周围。治疗２ 周,观察４ 周。结果　治疗组肿瘤生长较对照组缓慢,差异有显著性（ Ｐ＜ ０．０５） 。病理组织学检查可见治疗组肿瘤内片状及灶状坏死,肿瘤间质淋巴细胞浸润增多,肿瘤血管周围亦可见较多淋巴细胞聚集,脾小结明显增生,肝、脾及肺脏未见任何转移。对照组肿瘤细胞无明显坏死,间质淋巴细胞浸润很少,脾小结增生不明显,并观察到１ 例肺转移结节。结论　ＴＤＬＮＬ在体外经ＩＬ２ 短期活化后在肿瘤局部注射具有抗肿瘤作用,与以往经静     

Eosinophilic infiltration in advanced laryngeal squamous cell carcinoma

Tumor-associated tissue eosinophilia (TATE) has been related to prognosis in epithelial cancers, including cancers at several head and neck sites. This study prospectively examined 248 patients with stage in and IV laryngeal squamous cell carcinoma to determine prevalence and potential prognostic significance of TATE. Pretreatment tumor specimens were histopathologically evaluated. Presence and degree of TATE were analyzed with regard to other tumor characteristics, patient characteristics, and outcome criteria. Median follow-up was 48 months. Eosinophilia was found in 22.5% of specimens and was not related to tumor site, stage, patient age or sex, or treatment modality. Overall and disease-free survival rates and response to induction chemotherapy did not differ significantly with respect to TATE. This study represents the first long-term, prospective evaluation of TATE and its prognostic significance in a single head and neck site. Contrary to the findings of earlier preliminary reports, our results suggest that TATE is not a clinical useful prognostic parameter in advanced laryngeal squamous cell carcinoma.     

Studies on localization of laryngeal carcinoma associated antigen by immunoelectromicroscopy]

OBJECTIVE: To study the localization of laryngeal carcinoma associated antigen (LCAA) in the carcinoma tissue. METHODS: Ninety cases of laryngeal carcinoma and 14 cases of laryngeal precancerous lesion and 10 cases of normal laryngeal mucosa were detected with three strains of monoclonal antibodies LC9, LC11, LC12 by immunochemistry. The positive sections of laryngeal carcinoma were observed under light microscope and electromicroscopy. RESULTS: The positive rates of LCAA were dramatically higher than that in normal epithelial and precancerous tissues (P < 0.01). The results showed that the mixed monoclonal antibody had tissue specificity. The MLC associated antigens only distributed in cell membranes and/or cytoplasm. No cell nucleus was stained. CONCLUSION: The LCAA is mainly located in cell membranous structure. This study may provide morphological basis for immunoimaging diagnosis and targeting chemotherapy by application of laryngeal carcinoma McAb.     

局部复发喉癌的发生学研究

局部复发是喉癌的主要致死原因之一,保证安全切缘是防止喉癌术后复发的主要手段。但研究发现局部复发喉癌病例中27.8%切缘安全(>5mm,甚至>20mm),很难用残留肿瘤导致复发来解释[1]。如果原有器官内微小转移灶或是术中器官内种植,则复发肿瘤与原发肿瘤是相同克隆来源,通过遗传学检测应具同源性;如果切缘安全的病例是再发肿瘤,而非“复发”,与原发肿瘤克隆来源应不同。临床上对复发和再发的区别完全是靠时间来界定的,即>5年的复发肿瘤称为再发,<5年则为复发。显然用分子生物学方法更能精确区分这两个概念。2003—2004年对喉癌的局部复发灶与原…     

Relationship among lymphatic metastasis, pericancerous lymphocytic reaction and dendritic cell infiltration in laryngeal carcinoma cells]

OBJECTIVE: To study the role and the clinical significance of dendritic cell infiltration against tumor cells in patients with laryngeal squamous cell carcinoma. METHODS: Immunohistochemical method using S-100 protein antibody was employed to detect dendritic cells (DC) on paraffin-embedded tissue sections from 23 patients with laryngeal squamous cell carcinomas. The relationships among the density of dendritic cell infiltration in laryngeal carcinomas and cervical lymphatic metastasis, pericancerous lymphocytic reaction and other clinicopathologic parameters were observed. RESULTS: The numbers of dendritic cells infiltrating among laryngeal carcinoma cells in patients with no cervical lymph metastasis was significantly larger than those with cervical lymphatic metastasis (t = 4.889, P < 0.01). Significant increase DC infiltration among laryngeal carcinoma cells was found in the group with intensively positive reaction of pericancerous lymphocyte than in the medium and weakly positive groups. The number of infiltrating dendritic cells in patients who had survived more than 5 years was significantly larger than those survived less than 5 years (t = 4.423, P < 0.01). The numbers of the infiltrating dendritic cells in patients with well-differentiated squamous cell carcinoma and poorly differentiated squamous cells carcinoma were 2.2 +/- 1.07 and 14.6 +/- 7.14 respectively, which were significantly different (q = 4.532, P < 0.05). CONCLUSION: The study showed that DC infiltration among laryngeal squamous carcinoma cells played an important role in the host immune reaction against tumor. It indicated that when the density of infiltrating DC is higher, the patients had less chance of cervical lymph metastasis and may have a longer survival time. There was a coordinative effect between DC infiltration in tumor cells and pericancerous lymphocyte reaction. Thus, the DC infiltration among laryngeal squamous cells could be used as an index of prognosis.     

Infiltration of dendritic cells into cervical lymph nodes in laryngeal carcinoma]

OBJECTIVE: Dendritic cells(DC) form a system of highly efficient antigen-presenting cells. An immunohistochemical study was performed to prove their role in immune responses against cancer. METHODS: Paraffin blocks were prepared for staining with antibody against S-100 proteins in 157 lymph nodes obtained from elective cervical lymphadenectomy in 47 cases of laryngeal squamous cell carcinomas. RESULTS: The extent of infiltration by DC into the regional lymph nodes was evaluated in two groups according to negative and positive lymph node metastasis. Densities of DC ranged from 23.0 to 111.9, with an average of 57.2 +/- 16.0, in the negative metastasis nodes; and from 17.5 to 63.6, with an average of 38.3 +/- 14.9, in the positive metastasis nodes. The test revealed that the differences in population density between the two groups were significant (P < 0.001). Quantitative analysis showed that patients who survived longer than 5 years had significant higher numbers of DC in the lymph nodes than those who lived less than 5 years (P < 0.001). Densities of DC ranged from 32.0 to 111.9, with an average of 61.6 +/- 19.6, in the nodes of 20 cases of patients who survived longer than 5 years and from 17.5 to 65.8, with an average of 37.9 +/- 13.7, in the nodes of 27 cases of patients who survived less than 5 years. CONCLUSION: DC may be more directly involved in the host immune reaction against tumor by means of the most important antigen-presenting cells. DC in the cervical lymph nodes are important for establishing immunologic defense mechanisms in cases of laryngeal carcinomas, and the densities of DC can serve as a useful indicator in assessing prognosis of laryngeal carcinomas.     

Expression of p53 antigen in laryngeal carcinoma

The results of immunohistochemical investigations of p53 presence in 50 patients with laryngeal carcinoma were presented. In the whole investigated group the presence of p53 protein in 90% of all investigated cases was observed. In patients with advanced clinical stage of laryngeal carcinoma higher expression of p53 protein comparing with patients with lower clinical stages was more frequently observed--but it wasn't significant. No significant difference in presence of p53 protein among the patients with a different stage of histological differentiation of carcinoma and among the patients with present and absent metastatic changes in regional lymph nodes was observed.     

Expression of Bcl-2 antigen in laryngeal carcinoma

The results of immunohistochemical investigations of BCL-2 presence in 50 patients with laryngeal carcinoma were presented. In 14% of all investigated cases presence of BCL-2 protein was observed. In patients with advanced clinical stage of laryngeal carcinoma higher expression of BCL-2 protein comparing with patients with lower clinical stages was more frequently observed--but it wasn't significant. No significant difference in presence of BCL-2 protein among the patients with a different stage of histological differentiation of carcinoma and among the patients with present and absent metastatic changes in regional lymph nodes was observed.