ESR1基因单核苷酸多态性及单倍型与精神分裂症的关联研究

目的 探索雌激素受体1 (estrogen receptor 1,ESR1)基因rs2234693、rs9340799和rs3798759位点单核苷酸多念性(single nucleotide polymorphisms,SNPs)及其单倍型与精神分裂症(schizophrenia,SZ)发病之间的相关性.方法 应用聚合酶链反应-限制性片段长度多态性技术对333例SZ患者和315名正常对照rs2234693、rs9340799和rs3798759位点进行基因分型,应用x2检验对SZ组和对照组等位基因、基因型和单倍型频率进行分析.结果 rs2234693、rs9340799位点两组间基因型频率及等位基因分布差异均无统计学意义(P＞0.05).SZ组rs3798759位点GG基因型频率及G等位基因频率均高于健康对照组(P＜0.01).性别分层分析提示,女性SZ患者rs3798759位点TG、GG基因型频率及G等位基因频率均高于健康女性(P＜0.05).单倍型C-A-G和C-G-G在SZ组的分布频率高于对照组(P＜0.05).结论 rs3798759位点突变可能为女性精神分裂症发生的风险因子,C-A-G和C-G-G单倍型可能为精神分裂症的遗传风险单倍型.     

甲状腺乳头状癌中hMLH1、hMSH2和PCNA的表达及临床意义

目的观察hMLH1、hMSH2和PCNA在甲状腺乳头状癌中的表达及三者与肿瘤发生、发展的关系。方法应用免疫组化PV-6000两步法对甲状腺乳头状癌及癌旁正常甲状腺组织中hMLH1、hMSH2和PCNA的表达进行检测,并进行统计学分析。结果三种蛋白在甲状腺乳头状癌与癌旁正常组织中的阳性率差异有显著性(P<0.05),PCNA和hM-LH1蛋白与肿瘤包膜侵犯和淋巴结转移相关(P<0.05),hMSH2与淋巴结转移密切相关,但三者均与患者年龄、性别、肿瘤大小无相关性(P>0.05)。PCNA与hMSH2表达呈正相关(P<0.05)。结论 hMLH1、hMSH2和PCNA的异常表达与甲状腺乳头状癌的发生、发展密切相关。     

散发性结直肠癌hMLH1和hMSH2蛋白表达

目的　研究hMLH1及hMSH2两种错配修复 (mismatchrepair,MMR)蛋白在散发性结直肠癌中的表达变化并评估其可能的临床意义。方法　应用EnVision免疫组化两步法检测 1 1 1例散发性结直肠癌中hMLH1和hMSH2的蛋白表达变化 ,采用Kaplan Meier曲线、Log rank检验分析hMLH1蛋白表达变化与患者生存率之间的关系。 结果　 1 1 1例散发性结直肠癌中 ,hMLH1失表达有 1 9例 ,占 1 7 1 % (1 9/ 1 1 1 ) ,hMSH2失表达有 2例 ,占 1 8% (2 / 1 1 1 ) ,两者之和占总散发性结直肠癌病例的 1 8 9% (2 1 / 1 1 1 )。hMLH1或hMSH2蛋白失表达与患者肿瘤部位、组织学类型密切相关。近端结肠、低 -未分化腺癌及黏液腺癌中MMR异常表达比例高 (P <0 0 5 ) ,而与患者年龄、性别、肿瘤大体类型、肿块大小、浸润深度、淋巴结转移与否以及患者的Dukes分期均无显著性相关 (P >0 0 5 )。癌组织中hMLH1正常表达及失表达患者的 5年生存率分别为 6 9 5 7%及73 6 8% ,8年生存率分别为 5 3 5 8%及 73 6 8% ,8年生存率差别较明显 ,然差别无统计学显著性 (P >0 0 5 )。结论　一定比例的散发性结直肠癌中存在MMR基因的缺陷 ,其中hMLH1所起的作用远远大于hMSH2 ,hMLH1失表达与否可能成为有意义的远期生存预后指标     

新疆地区汉族和维吾尔族部分人群HLA-DRB1、HLA-DQB1、HLA-DPB1基因多态性与单倍型的比较

目的了解中国新疆维吾尔自治区汉族和维吾尔族人类白细胞抗原DRB1(HLA-DRB1)、HLA-DQB1、HLA-DPB1高分辨分型基础上的基因多态性和单倍型分布的特点。方法应用聚合酶链反应-直接测序基因分型(PCR-SBT)法对新疆地区188例汉族人群与90例维吾尔族人群进行HLA-DRB1、HLA-DQB1、HLA-DPB1基因分型,直接计数法计算基因频率,应用Arlequin3.5软件以最大似然法获得单倍型频率。结果汉族人群频率最高的HLA-Ⅱ类基因分别是DRB1*09∶01(12.8%)、DQB1*03∶01(20.3%)和DPB1*05∶01(33.5%);三位点组成的单倍型频率最高的分别是DRB1*09∶01-DQB1*03∶03(8.8%)、DRB1*09∶01-DPB1*05∶01(5.0%)、DQB1*03∶01-DPB1*05∶01(9.6%)和DRB1*09∶01-DQB1*03∶03-DPB1*05∶01(6.3%)。维吾尔族人群频率最高的HLA-Ⅱ类基因分别是DRB1*07∶01(18.3%)、DQB1*03∶01(18.9%)和DPB1*04∶01(30.0%);三位点组成的单倍型频率最高的分别是DRB1*07∶01-DQB1*02∶02(16.1%)、DRB1*07∶01-DPB1*04∶01(5.0%)、DQB1*03∶01-DPB1*04∶01(7.3%)、DRB1*07∶01-DQB1*02∶02-DPB1*04∶01(7.4%)。两组人群中等位基因有显著差异的是∶DRB1*04∶04、DRB1*07∶01、DRB1*08∶03、DRB1*11∶04、DRB1*12∶02、DRB1*13∶01、DRB1*15∶01、DQB1*02∶02、DQB1*06∶01、DPB1*04∶01和DPB1*05∶01。结论中国新疆汉族与维吾尔族人群HLA-DRB1、HLA-DQB1、HLA-DPB1等位基因频率、单倍型频率分布均有自己的多态性特征。     

基于标签单核苷酸多态性单倍型和单倍域的构建及其在关联研究中的应用

人类基因组中单倍型(haplotype)和单倍域(haplotype block)的结构提供了人类进化的宝贵信息,并成为发现人类复杂疾病易感基因的有效策略。一个单倍域可分割成多个具有有限单倍型多样性的离散的区域,代表每个区域结构特征的少量标签单核苷酸多态性(tag single nucleotide polymorphism,tSNP)可使绝大部分单倍型相互区分开来。因此,标签SNP在单倍型和单倍域的构建和关联研究中具有重要地位。构建单倍型和单倍域的方法分为两类,分别是基于大家系中基因分型数据和基于统计学的算法。通过系统回顾几种单倍型和单倍域的构建方法,了解它们在不同的疾病模型或根据不同的分割标准,进行关联研究的检验效能,客观评价每种方法的优、缺点、应用前景及其在关联研究中的应用。随着国际人类基因组单倍型图的完成和单倍型构建统计学运算规则的完善,融合数学、物理学、计算机科学等学科的单倍型构建方法将对人类遗传学、复杂疾病易感基因的定位和克隆鉴定等生命科学的相关领域产生深远的影响。     

浙江地区汉族人群caspase-3基因三个位点的单倍型研究

目的分析汉族人群caspase-3基因的单核苷酸多态性(singlenucleotidepolymorphisms,SNPs)位点及其构成的单倍型,为研究caspase-3基因对凋亡调节机制的个体差异提供线索。方法用变性高效液相色谱技术和DNA测序技术检测caspase-3基因的调控区、第2～7外显子及部分侧翼序列的多态性位点;分析位点间的连锁不平衡关系,估算它们构成的单倍型。结果共检出3个SNP位点(C829A、A17532C、C20541T),分别位于caspase-3基因的5′端调控区、第4内含子和3′调控区;3个位点间存在强连锁不平衡,其中位点A17532C与C20541T呈完全连锁不平衡;54.3%的C-829/A-17532/C-20541是汉族人群的主要单倍型。结论浙江地区汉族人群caspase-3基因上的3个SNP位点间存在强连锁不平衡,它们构成的主要单倍型不同于北美人群。     

湖北土家族人群HLA-A、B等位基因及单倍型多态性的分布

目的研究湖北五峰县土家族190名无亲缘关系健康个体HLA-A、B等位基因及单倍型频率的分布，为进一步研究HLA基因多态性与疾病的关联奠定基础。方法采用序列分型（sequence-baaed typing，SBT）法对最具多态性的HIA-A、B基因2、3外显子进行了基因型分析，采用Arlequin软件对群体基因、单倍型频率进行最大估计值的计算。结果HLA-A、B等位基因符合Hardy-Weinberg平衡定律（P〉0．05），无显著的连锁不平衡现象，共检测出26个HLA-A等位基因及41个HLA-B等位基因，其中频率最高的为A＊0201（0．16053），A＊110101（0．14737），A＊24020101（0．14211），B＊4001（0．14737），B＊4601（0．13947），其次为A＊0207（0．08947），A＊0206（0．08158），B＊1301（0．07632），B＊5801（0．08947），B＊1501（0．09737），而大于0．05％的为A＊330301（0．05526），B＊1502（0．05526），B＊3501（0．05263），单倍型分布较普遍的为A＊0202-B＊4001（0．04196），A＊0201-B＊4601（0．03625）。结论采用高分辨测序分型法对土家族人群HLA-A、B等位基因进行分型的实验结果可做为湖北土家族人HLA-A、B等位基因、单倍型频率的群体资料，对进一步开展群体遗传学、临床器官移植、疾病关联、HLA遗传学特征、法医学、人类学等研究具有重要意义。     

广东汉族人群HLA Ⅰ、Ⅱ基因多态性及单倍型分析

目的 检测广东汉族人群HLA-A、B、Cw、DRB1基因频率，分析该人群HLAⅠ、Ⅱ等位基因多态性及其单倍型特点。方法 骨髓移植供者160人，抗凝血提取DNA，半量全自动聚合酶链反应-单链构象多态分型检测HLA-A、B、Cw、DRB1基因型。结果 在低分辨水平分别检出HLA-A、B、Cw及DRB1等位基因12、23、11、13个。统计分析呈现显著连锁不平衡的HLA-A-B单倍型9个，B-Cw单倍型20个，A-Cw单倍型7个，HLA-A-DRB1单倍型8个，B-DRB1单倍型9个，Cw-DRB1单倍型10个。结论 广东汉族群体HLA-基因具有较为丰富的多态性，其双座位连锁不平衡单倍型具有地区性遗传特征。     

慢性乙型肝炎转化生长因子β1基因单核苷酸多态性及单倍型分析

目的探讨转化生长因子β1（TGF-β1）基因单核苷酸多态性（SNP）及其单倍型与慢性乙型肝炎易感性之间的关系。方法以155例慢性乙型肝炎患者和170例健康对照者为研究对象，应用聚合酶链反应．限制性片段长度多态性（PCR-RFLP）和DNA测序的方法对TGF-β1基因-509C／T、869T／C（Leu10Pro）单核苷酸多态性进行基因分型，同时采用酶联免疫吸附试验（ELISA）检测血清TGF-β1水平。用SHEsis软件分析TGF-β1基因的连锁不平衡及单倍型频率。结果TGF-β1基因869T／C（Leu10Pro）多态性在慢性乙型肝炎组和正常人群中的分布差异无统计学意义（P〉0．05），而TGF-β1基因-509C／T多态性在两组人群中的分布差异有统计学意义（P〈0．05），等位基因频率的相对风险分析发现，T等位基因携带者患慢性乙型肝炎的风险是C等位基因的1．507倍（OR=1．507，95％CI：1．106～2．054）；携带T等位基因的慢性乙型肝炎患者血清TGF-β1水平显著高于不携带者（P〈0．05）。联合基因型分析发现，TGF-β1基因-509C／T、869T／C单核苷酸多态性存在着强烈的连锁不平衡（ID’I=0．891），与-509C／869T单倍型携带者比较，-509T／869C单倍型携带者显著增加了慢性乙型肝炎的发病风险（OR=1．663，95％CI：1．193-2．320）。结论TGF-β1基因-509C／T多态性和-509T／869C单倍型与慢性乙型肝炎的发病具有相关性，其中T等位基因可能是慢性乙型肝炎的遗传易感基因，携带T等位基因的个体可能通过促进TGF-β1的高度表达进而增加了慢性乙型肝炎的发病风险。     

NURR1基因多态性与帕金森病的关联研究

目的 了解NURR1基因多态性与四川地区散发性帕金森病之间的相关性.方法 采用病例-对照研究,应用聚合酶链反应、等位基因特异性、限制性片段长度多态性对四川地区汉族人群241例帕金森病患者和236名正常对照NURR1基冈启动子区的c.-2922(C)2-3及第6内含子的ⅣS6+18imG多态位点进行关联分析.结果 IVS6+18insG位点帕金森病组3G/3G,3G/2G,2G/2G基因型频率与对照组相比差异无统计学意义(X2=3.733,P=0.155).进一步按发病年龄分层后发现,50岁以前发病的帕金森病患者基因型频率与对照组之间差异有统计学意义(X2=6.545,P=0.038).发病年龄＜50岁的帕金森病组患者3G/2G基因型频率显著高于对照组(54.12%vs 38.14%),并且与其他两组基因型合并相比差异有统计学意义(X2=6.537,P=0.011;OR=1.913,95%CI:1.159～3.158).c.-2922(C)2-3位点帕金森病组与对照组相比3C/3C,3C/2C及2C/2C基因型频率差异无统计学意义(P=0.766).结论 本研究结果提示NURR1基因ⅣS6+18insG多态可能与本组人群早发性帕金森病的遗传易感性相关;未发现c.-2922(C)2-3位点多态性与本组人群帕金森病的遗传易感性相关.     

Association of genetic polymorphisms and haplotypes in hMLH1 and hMSH3 gene with the risk of papillary thyroid carcinoma

OBJECTIVE: To explore the relationship of the genetic polymorphisms and the haplotypes in hMLH1 and hMSH3 gene with the risk of papillary thyroid carcinoma (PTC) in Chinese Hans. METHODS: A hospital based 1:1 matched case-control study was carried out. The polymorphisms for 204 pairs of PTC cases and healthy controls were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and allele specific oligonucleotide (PCR-ASO) assays. RESULTS: (1) The PTC risk was marginally increased in the hMLH1 1151TA genotype, with odds ratio (OR) of 2.15 (95%CI: 0.99-4.85); the PTC risk was significantly increased in the mutant genotype 1151TA+AA, with OR of 2.15 (95%CI: 1.02-4.69); (2) The haplotypes of -93G, 1151A, 655A in the hMLH1 gene could increase the PTC risk, with OR of 2.67 (95%CI: 1.16-6.53, P=0.011), compared with the haplotype of -93G, 1151T, 655A; (3) Compared to 3124A, 2835G haplotype in hMSH3 gene, the 3124G, 2835A haplotype could increase the PTC risk marginally, with OR of 3.08 (95%CI: 0.92-13.25). CONCLUSION: The 1151T/A polymorphism in hMLH1 was associated with PTC; both the haplotype of -93G, 1151A, 655A in hMLH1 and the 3124G, 2835A haplotype in hMSH3 were associated with PTC.     

Single nucleotide polymorphisms of ataxia telangiectasia mutated and the risk of papillary thyroid carcinoma

Genetic factors associated with susceptibility to papillary thyroid carcinoma (PTC) are not well known. We evaluated the association between single nucleotide polymorphisms (SNPs) of ataxia telangiectasia mutated (ATM) and the risk of PTC. A total of 437 histologically confirmed PTC cases and 184 cancer‐free controls without thyroid nodules were recruited. Genotypes with respect to five ATM SNPs (rs189037, rs664677, rs373759, rs664143, and rs4585) were determined by the TaqMan assay, and odds ratios and 95% confidence intervals were obtained by logistic regression analysis. Linkage disequilibria and haplotypes were examined from the genotype data. When evaluated separately the genotype distributions of the five ATM SNPs were similar in the PTC cases and controls. Three ATM SNPs (rs373759, rs664143, and rs4585) were found to be in strong linkage disequilibrium (D′ = 1.00, P < 0.001). When the three haplotypes (C‐A‐G), (T‐G‐T), and (C‐G‐T) of these three ATM SNP sites were analyzed, ATM haplotype (C‐G‐T) +/? was associated with a lower risk of PTC than ATM haplotype (C‐G‐T) ?/? (P = 0.03) after adjusting for age and gender. Our results suggest that genetic polymorphisms of ATM may play an important role in the development of thyroid cancer in the Korean population. Environ. Mol. Mutagen. 56:70–76, 2015. © 2014 Wiley Periodicals, Inc.     

Distinction between papillary thyroid hyperplasia and papillary thyroid carcinoma by immunohistochemical staining for cytokeratin 19, galectin-3, and HBME-1

The histopathology of papillary thyroid hyperplasia and papillary thyroid carcinoma is similar enough to cause a diagnostic dilemma in a few cases. Both lesions may have papillary fronds with fibrovascular cores, nuclear crowding, and nuclear anisocytosis. Formalin-fixed paraffin-embedded tissues from 30 randomly selected patients with papillary thyroid hyperplasia and an equal number from patients with papillary thyroid carcinoma were analyzed for expression of cytokeratin 19 (CK19), galectin-3, and HBME-1. Cases of papillary thyroid carcinoma had moderate to strong CK19, galectin-3, and HBME-1 reactivity although both CK19 and galectin-3 showed positive staining in a significant number of nonneoplastic thyroid cases. HBME-1 was uncommon in the nonneoplastic cases. These results indicate that HBME-1 may be useful in helping to distinguish papillary thyroid carcinoma from hyperplasia in diagnostically difficult cases.     

Galectin-3 expression in papillary microcarcinoma of the thyroid

AIMS: Galectin-3 is a beta-galactoside binding protein, recently recognized as a promising molecular marker of thyroid malignancy. As reported in several studies, galectin-3 is highly expressed in papillary thyroid carcinoma, but its expression has not been investigated in papillary microcarcinoma, which is a variant of papillary thyroid carcinoma. METHODS AND RESULTS: Using a monoclonal antibody to galectin-3 and the avidin-biotin-peroxidase complex (ABC) immunohistochemical technique, we analysed galectin-3 expression in 63 cases of papillary microcarcinoma. The results showed immunohistochemical reactivity for galectin-3 in 51 (80.9%) cases. Intensity of staining varied from strong or moderate to weak. Galectin-3 localization was mostly cytoplasmic, but also membranous or nuclear in some cells. Immunohistochemical expression of galectin-3 was not found in 12 (19.1%) cases. Most galectin-3 negative microcarcinomas (10/12) were of the non-classical type, i.e. without papillary architecture. Neither the frequency nor the intensity of a positive reaction was related to tumour size. CONCLUSIONS: Galectin-3 gene is expressed at the protein level in most papillary microcarcinomas, although with slightly lower frequency than that reported for clinically evident papillary thyroid carcinoma. The presence of galectin-3 in clinically silent microcarcinomas may indicate that galectin-3 is not related to growth or aggressiveness of papillary thyroid microcarcinomas but rather plays some other role in thyroid tumour biology.     

甲状腺乳头状癌BRAF基因突变的检测及临床意义

目的 了解甲状腺乳头状癌(papillary thyroid carcinoma,PTC)BRAF基因T1799A点突变的情况与临床病理学特征的关系.方法 应用聚合酶链反应及DNA直接测序法对43例PTC患者,20例非PTC甲状腺病变患者及40份正常甲状腺组织对照的新鲜标本进行BRAF基因检测.分析BRAF基因突变与性别、发病年龄、原发灶大小、甲状腺包膜外浸润、颈淋巴结转移及远处转移等临床病理学特征的关系.结果 43例PTC中1 7例检出BRAF基因T1799A点突变.检出率为39.5%,而在非PTC甲状腺病变患者和40份正常甲状腺组织未发现T1799A点突变.BRAF基因突变与PTC甲状腺包膜外浸润及颈淋巴结转移密切相关(P<0.05及P<0.05),与性别、发病年龄、原发灶大小及远处转移无关.结论 BRAF基因突变与颈淋巴结转移和甲状腺包膜外浸润密切相关,突变可能增加PTC的侵袭性并影响预后.     

Pitfalls in the surgical pathologic diagnosis of papillary thyroid carcinoma

While the surgical pathology examination of the thyroid gland for papillary carcinoma may seem, on its surface, to be relatively straightforward, in reality it is fraught with diagnostic traps. Avoidance of these pitfalls is necessary for guiding the surgeon and endocrinologist to the appropriate treatment and follow up. This review will detail a selected group of some of the more commonly encountered challenges in making the diagnosis of papillary thyroid carcinoma from a busy head and neck pathology consultation practice.     

Synchronous papillary thyroid carcinoma and follicular thyroid carcinoma: case report and review of literature

Collision tumor is a term denoting two histologically distinct tumor types occuring at the same anatomic site, which is a rare clinical entity. In the thyroid gland, collision tumors are rare. Here we report a case of the synchronous occurrence of follicular thyroid carcinoma (FTC) and papillary thyroid carcinoma (PTC). The current case report describes a 40-year-old woman with synchronous FTC and PTC. Pathologists and surgeons should be aware of collision tumors to avoid possible misdiagnosis.     

Coexistence of primary squamous cell carcinoma of thyroid with classic papillary thyroid carcinoma

Primary squamous cell carcinoma of the thyroid gland is very rare and its histogenesis is poorly defined so far. Although there have been some cases of squamous cell carcinoma with variant types of papillary thyroid carcinoma (PTC), the present case is the first primary squamous cell carcinoma with classic PTC to be reported. A 43‐year‐old woman presented with a 20 year history of neck mass. Neck ultrasound indicated a 6 × 4 × 3 cm large mass. The patient underwent total thyroidectomy. Histopathology indicated a well‐differentiated squamous cell carcinoma and squamous metaplasia in conjunction with classic PTC. On immunohistochemistry cytokeratin 7 was positive in papillary carcinoma and squamous metaplasia, thyroglobulin was positive only in papillary carcinoma, and p63 was positive in squamous metaplasia and squamous cell carcinoma. Postoperatively, the patient received 59.4 Gy adjuvant radiotherapy, hormonal therapy and radioactive iodine therapy. At 8 months after surgery the patient remained disease free.