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1、 骨纤维结构不良病变组织雌激素受体和增殖细胞核抗原检测及临床意义 作者：牛彦辉，王臻，郑修存，吴作栋，沈强，张立明【关键词】 ,纤维发育不良 Detection and significance of estrogen receptor and proliferating cell nuclear antigen in tissues of fibrous dysplasia of bone【Abstract】 AI
2、M: To investigate the relation between the clinical biological behavior and the expression of estrogen receptor (ER) and proliferating cell nuclear antigen (PCNA) in the tissues of fibrous dysplasia of bone (FDB). METHODS: Ninetyeight cases wrer surgically treated, pathologic
3、ally confirmed as having fibrous dysplasia and followed up. The primary operation specimens were randomly selected and divided into recurrence group (n=24) and cured group (n=24), children group (n=26) and adult group(n=22). The ER and PCNA in the specimens were detected with monoclonal antibo
4、dy to ER or PCNA by immunohistochemistry. RESULTS: The general positive expression rate of ER was 56.25%(27/48). The positivity rate in recurrence group was higher than that in cured group (70.83% vs 41.67%, P0.05) and the positivity rate in children group was higher than that in adult group (
5、69.23% vs 40.91%, P0.05). The expression of PCNA in recurrence group was higher than that in cured group (P0.01). The expression of PCNA had a tendency to be higher in children group than that in adult group, but with no statistical significance (P0.05). CONCLUSION: FDB is relative to the estr
6、ogen. The abnormal expression of ER, the disturbance of the regulation of estrogen in bone development and metabolism may be the causes of the onset and recurrence of FDB. The expression of PCNA is related with the recurrence of FDB. PCNA can be used as a reference marker for prognosis of FDB.【Keywo
7、rds】 fibrous dysplasia of bone; receptors estrogen; proliferating cell nuclear antigen(PCNA); immunohistochemistry【摘要】 目的： 探讨骨纤维结构不良（FDB）病变组织雌激素受体（ER）和增殖细胞核抗原（PCNA）的表达并讨论其临床意义. 方法： 98例FDB患者手术治疗、病理确诊并获随访. 收集初次手术切除的标本，随机从治愈和复发病例中各抽取24例. 对复发组（n=24）与治愈组（n=24）、小儿组（n=26
8、）与成人组（n=22）进行对比，用单克隆抗体免疫组织化学方法对病变组织进行ER和PCNA检测. 结果： ER总阳性率为56.25%(27/48). 复发组70.83%（17/24）明显高于治愈组41.67%(10/24) （P0.05）. 小儿组69.23%(18/26)明显高于成人组40.91%(9/22， P0.05). 复发组PCNA阳性表达强度显著高于治愈组(P0.01)，而小儿组PCNA阳性表达高于成人组，但无统计学差异(P0.05). 结论： FDB的雌激素相关性表现为ER异常表达而导致骨发育及代谢调节紊乱；PCNA阳性表达强度与骨纤维结构不良的复发密切相关，可作为其预后估测的参考
9、指标.【关键词】 纤维发育不良，骨；受体，雌激素；增殖细胞核抗原；免疫组织化学0引言骨纤维结构不良（fibrous dysplasia of bone, FDB）的发病率居骨与关节瘤样病变的首位1，其临床表现为，术后复发率高，儿童期更具高度复发倾向和侵袭性，部分病灶呈进展性，部分病灶却长期静止，有些病例反复多次复发，少数可发生恶变，但泛发性骨纤维结构不良（Albright综合征）主要见于女性. 我们检测骨纤病变组织中雌激素受（estrogen receptor, ER）和增殖细胞核抗原（proliferating cell nuclear antigen, PCNA）的变化并讨论其
10、临床意义.1材料和方法1.1材料标本来源于197701/199806西京医院骨科手术治疗的骨纤患者病变组织石蜡埋块. 选取其中有随访结果的98例患者初次手术标本，根据随访结果分成复发与治愈两组. 采用随机数字法各选取24个标本，共48例. 再根据患者年龄分为：小儿组（14岁）26例，成人组（115岁）22例. 一抗分别选用法国Immunotech公司产的鼠抗人ER mAb（150）和丹麦DAKO公司产的鼠抗人PCNA mAb(140). 二抗均选用Sigma公司产的生物素标记羊抗鼠IgG. SABC试剂盒(武汉博士德生物工程有限公司).1.2方法标本连续切片3张，1张HE染色，病理医师再次确诊
11、，2张分别进行ER和PCNA检测. 检测采用免疫组织化学染色法（SABC法）. 步骤： 切片脱蜡至水；6 mL/L H2O2封闭内源性过氧化物酶，抗原修复10 min; 20 mL/L羊血清封闭20 min；加一抗4冰箱过夜；37水浴箱孵育1 h；滴加二抗37 20 min. 以上三步间均用PBS缓冲液振洗5 min3次；新配DAB显色液显色；苏木素衬染，乙醇脱水，透明，封片. ER染色选用ER阳性乳腺癌组织切片为阳性对照，PCNA染色选用PCNA阳性扁桃体增生组织切片为阳性对照. 均以PBS缓冲液代替一抗做阴性对照.用Olympus1双目显微镜观察细胞. ER染色： 胞质和胞核内可见棕黄色颗
12、粒者为阳性细胞. 每片随机观察10个高倍视野，平均阳性细胞数>5%定为ER阳性. PCNA染色： 细胞核内显示棕黄色颗粒者为阳性细胞. 随机观察5个高倍视野，每视野计数100个细胞中的阳性细胞数，求均数. 采用半定量法，其阳性表达强度为，: 5%; +: 5%15%; : 15%30%; : 30%. 统计学处理： ER阳性率组间比较采用2检验，PCNA阳性表达强度组间比较采用Wilcoxon等级秩和检验.2结果 2.1ER和PCNA阳性表达时细胞中的定位及组织中阳性细胞分布规律ER阳性表达时，阳性细胞胞质及胞核中均可见棕黄色阳性颗粒，分布均匀（Fig 1）. 骨小梁
13、周围的骨母细胞及胞体较宽大的纤维样细胞表达强（Fig 2），瘦长的纤维样细胞表达较弱. PCNA阳性表达时，仅胞核内可见棕黄色阳性颗粒，分布不均，阳性胞核着色深浅不一. 骨母细胞及胞体较宽大的纤维样细胞多呈阳性，瘦长的纤维样细胞多为阴性（Fig 3）. 组织中阳性细胞分布不均，纤维样细胞宽大的部位阳性表达强，细胞瘦长的部位阳性表达弱（Fig 4） .2.2ER阳性表达结果48例中ER阳性率为56.25%（27/48）. 复发组阳性表达率70.83%(17/24)明显高于治愈组41.67%(10/24, P0.05)，而小儿组阳性表达率69.23%(18/26)明显高于成人组40.91%(9/2
14、2, P0.05, Tab 1).2.3PCNA阳性表达结果复发组PCNA阳性表达强度明显高于治愈组(P0.01)，而小儿组阳性表达高于成人组(P0.05, Tab 2). 表2各组PCNA阳性表达结果（略）3讨论 FDB是一种以骨纤维变性为特征的类肿瘤疾患，约占类肿瘤疾病的7%2. 疾病复发时，多以纤维组织增生为主，多为胞体及胞核较宽大的细胞，排列较致密，骨小梁数量少，短小或细长. 疾病治愈时纤维样细胞多细长，纤维组织排列较疏松，骨小梁相对粗大，数量也较多，甚至部分可连接成网. 免疫组化染色也显示：标本中宽大的纤维样细胞ER和PCNA的阳性表达较强，而以细长纤维样细胞为
15、主的标本中阳性表达较弱，甚至无阳性表达. 提示骨纤病变组织的形态学特征及ER和PCNA的阳性表达强度与其临床生物学行为密切相关. 纤维组织增生明显，ER及PCNA阳性表达强的病例，临床上易复发.ER是一种存在于某些组织细胞胞质和胞核中的蛋白质3. 雌激素通过骨组织中的ER发挥其调节作用4. FDB复发患者和小儿ER阳性率均明显高于治愈患者和成人(P0.05)，说明ER与骨纤维化密切相关. 正常骨组织中ER较少，ER阳性率较低，约25%4. 本组ER总阳性率为56.25%，明显高于正常骨组织. 提示骨纤具有雌激素相关性，ER异常表达，雌激素对骨发育及代谢调节紊乱可能是其发病和复发的原因之一.PC
16、NA是仅在增殖细胞胞核中合成与表达的一种核蛋白. 其阳性表达的高低与细胞增殖活性密切相关5. PCNA检测目前已广泛应用于肿瘤的临床及实验研究，对癌前病变的检测、良恶性肿瘤的鉴别、肿瘤的分级和预后判断等有重要价值. 骨纤的病理基础为纤维组织异常增生，组织学观察发现不同病例中纤维组织的增生程度各异. 临床上有些病灶呈进展性，而有些病灶却相对静止，有些病例反复复发. 有报道0.4%0.5%发生恶变6. 提示病变组织的增生活性存在差异，复发病例病变组织增生活跃，其PCNA表达强弱与骨纤的复发密切相关，是其预后估测的参考指标.【参考文献】1 冯乃实，李瑞宗，张学军，等. 骨与关节肿瘤及瘤样
17、病变4327例统计分析J. 中华骨科杂志，1997;17(2):760-763.Feng NS, Li RZ, Zhang XJ, et al. Statistics and analysis of 4327 cases of tumors and tumorlike lesions in bone and joint J. Chin J Orthop, 1997;17(2):760-763.2 Chow LT, Griffith J, Chow WH, et al. Monostotic fibrous dysplasia of the spine: Report of a ca
18、se involving the lumbar transverse process and review of the literature J. Arch Orthop Trauma Surg, 2000;120(78):460-464.3 傅红，莫善兢，朱慰祺. 雌孕激素受体检测及其在临床肿瘤研究中的应用J. 实用癌症杂志，1991;6(2):147-149.Fu H, Mo SJ, Zhu WQ. Application of the detection of estrogen receptor and pregnant receptor in clinical cancer rese
19、arch J. Prac J Cancer, 1991;6(2):147-149.4 韩守威，何斌，唐茜萍. 正常骨和骨肿瘤组织雌激素受体分析J. 华西医科大学学报，1993;24(2):160-163. Han SW, He B, Tang QP. Analysis of the estrogen receptors in the tissue of bone and in tumors of bone J. J West China Univ Med Sci, 1993;24(2):160-163.5 杨长春，姜鸿彦，黄维国. P
20、CNA与肿瘤J. 医学信息，2000;13(7):413. Yang CC, Jiang HY, Huang WG. PCNA and cancer J. Med Message, 2000;13(7):413.6 LopezBen R, Pitt MJ, Jaffe KA, et al. Osteosarcoma in a patient with McCuneAlbright syndrome and Mazabrauds syndrome J. Skeletal Radiol, 1999;28(9):522-526./Porganizati
21、on system and working mode, the selection of a number of municipal strengthening group to build innovation and strengthen the organization of the basic level organizations advanced model.(seven) the manual work of the Communist Youth League Organization published and League branch of manual work, to
22、 promote the use of two manuals in the citys organizations at various levels. The county committee, directly under the group (work) is under the jurisdiction of grassroots League and group (total) using branch ratio shall not be less than 70%.Three, to further strengthen the groups leadership and te
23、am building(a) to strengthen the leagues leadership at all levels of construction. Continue to do a good job of municipal Party Committee Organization Department on the strengthening of the Communist Youth League cadres management opinions implementation. To promote the group counties (autonomous co
24、unties, cities) Commission and city directly under the group (workers) appointed to carry out the Five team building activities. Improve the League Association Department working mechanism. Conscientiously fulfill the duties of CO, combined with the city group directly under (the) Committee of the g
25、eneral election and the adjustment, actively jointly with the unit where the party organization department good group leadership selection with the work. Do a good job in municipal Party committee assistant committee leadership and leading cadres of the annual assessment work, adhere to and improve
26、the assistant secretary of the Communist Youth League talk system group To weave individual conversation and collective conversation, to strengthen the committees of all levels, the Standing Committee of the Standing Committee, the establishment of a sound system of central group learning, the syste
27、m of investigation and study, the system of important issues and the reporting system.(two) to strengthen the work of education and training of cadres to conscientiously implement the Central Committee. On the implementation of grassroots cadres training project and opinions on carrying out the miss
28、ion municipal CYL Cadres training project implementation opinions, full implementation of the project of the City Youth League cadres training education. Focus on the county grassroots cadres and workers focused on training, the new school system vocational training League cadres, for the Municipal
29、Organization Department jointly held the third session of the Communist Youth League system training cadres in party. Focusing on the construction of new socialist countryside, strengthening village cadres and rural construction team group member of the young team, the county organizations at variou
30、s levels throughout the year is expected to train 100 00 rural grassroots cadres, plans to train 1000 village secretary of the village Party branch.(three) actively promote grassroots cadres election work. Under the unified leadership of the party organization, to strengthen grassroots organizations
31、 direct work, standardize the election work rules and procedures, in the designated candidate, open competition to democratically elected branch secretary at the same time, actively and steadily the development of Township, streets and other grassroots organizations in the direct election of the pil
32、ot work, on the basis of summarizing the experience and gradually expand the scope of the pilot, and gradually formed a set of measures and cadres election system.(four) to strengthen the cadres testing exercise. To study the development of XX city from 2006 to 2008 cadres testing exercise planning.
33、 To select outstanding cadres to the central mission in Beijing, Shanghai, Guangzhou and other central organs and sending work in developed areas. With the Central Committee the implementation of the western region and ethnic minorities cadres training plan . To carry out the eighth batch of municip
34、al organs attachment cadre management, selection, training work. The outstanding cadres to start the National Development Bank XX branch testing exercise.(five) to strengthen the group construction. The main position to play the role of Youth League cadre education and training as the city, in accor
35、dance with the requirements of post training, further curriculum scientifically and rationally, adopt arrangement menu course, basic theory includes group work, focus, hot and difficult issues, to enhance the content and practice; to strengthen the construction of teaching staff, the integration of
36、teachers, establish a quality teacher training team, establish and implement course evaluation system; take delivery of training to the countryside, on-site training, and increase the rural grassroots cadres training efforts.Four, efforts to strengthen the work of young talent(a) to tenth XX young 5
37、4 medal in recognition of the publicity, and actively explore the XX youth service. 54 Medal of institutionalization, standardization and socialization mechanism. Give full play to the XX award in recognition of 54 youth medal Association, Youth Award winner in the 54 chapter, investigation activiti
38、es. Who won the Youth Service medal, dedication to the society to create conditions, build the stage. During the 54, named in recognition of the tenth XX young 54 medal. The association of XX editing of young talent , the exchange of information and communication platform for the XX of outstanding y
39、oung talents.(two) the establishment of municipal Party committee of young talent information database. In accordance with the feasible, effective, classification management, implement step by step principle, the establishment of municipal Party committee of youth talent database; guiding organizati
40、ons at various levels generally established the young talent database, construction of city, county (directly under the Communist Youth League), grassroots organizations three young talent information database the relevant information collection system, and gradually establish the citys young people
41、, the Communist Youth League youth talent information network, provide information platform for young talent resource development.Five, take effective measures to strengthen the mission management(a) to strengthen the mission management work. Strengthen the dues collection and management, continue t
42、o adhere to the dues collection regularly reporting system, the annual comprehensive award in recognition of the tour and the tour one vote veto system audit system. Do statistical work in the group. The group service award in recognition of the advanced units, standardization, promoting group manag
43、ement work institutionalized.(two) to strengthen the organization of research and information exchange efforts. Grassroots carry out research, efforts to form a number of theoretical results. Make full use of the network carrier, to explore the establishment of electronic group service system, promo
44、te the work of Communist Youth League information.Attachment 4:XX municipal Party Committee Propaganda Department of the Communist Youth League work pointsIn 2006, the Communist Youth League propaganda ideological work XX to Deng Xiaoping theory and the important thought of Three Represents as guida
45、nce, study and implement the partys sixteen and the spirit of the the fifth Plenary Session of the 16th CPC Central Committee implementation of the eight plenary session of the two committee, the citys cultural work conference, the Propaganda Department of the meeting and the the fourth Plenary Sess
46、ion of the 15th CPC Central Committee permanent mission spirit, closely around 11th Five-Year XX the goal of full implementation of Scientific Outlook on Development, emancipating the mind, seeking truth from facts, keep pace with the times, close to reality, close to life, close to the young, perse
47、vere grasp orientation, grasping deepening, grasping construction innovation, further improve the groups propaganda and ideological work ability, unity To guide the majority of members of the youth to achieve the 11th Five-Year planning tasks, the overall construction of a well-off society, building
48、 a socialist harmonious society to make new contributions.One, adhere to the Three Represents important thought and Scientific Outlook on Development education to guide the youth, to further strengthen the ideological and political education of youth(a) armed with youth science theory. Unremittingly with Deng Xiaoping theory and the important thought of Three Represents armed members of youth education, to promote the study and implement to expand the depth and breadth. To study and implement the important thought of Thr
