韧带样型纤维瘤病的临床病理学及遗传学研究

目的研究韧带样型纤维瘤病的临床病理学和遗传学特点,探讨用荧光原位杂交的方法在石蜡包埋组织中检测8号染色体三体的可行性。方法分析96例韧带样型纤维瘤病的临床资料,对69例进行组织学形态和免疫学表型分析（免疫组织化学EnVision两步法,抗体包括波形蛋白、结蛋白、α-平滑肌肌动蛋白、B—catenin、CD34、CD117、S-100）、对2例行电镜观察,采用荧光原位杂交检测20例石蜡包埋组织中的8号染色体三体。结果96例病例中男20例,女76例,年龄范围8～86岁,平均35．3岁。腹部以外部位的病变44例,腹壁和盆腔的病变28例,腹腔内病变23例,其中2例与Gardner综合征伴发。病变直径在0．6～24．0cm之间,平均8．4cm,结节或者条索状肿块,质地韧,切面粗糙。显微镜下形态较一致的梭形细胞分布于大量增生的胶原纤维间质内,梭形的纤维母细胞和肌纤维母细胞常排列成束状,胞质界限不清,细胞异形性不明显。透射电镜下,纤维母细胞有丰富的粗面内质网和发达的高尔基复合体,肌纤维母细胞还可以看到应力纤维、纤维连接复合体、中间连接和缝隙连接。所有病例均表达波形蛋白,部分病例还表达结蛋白和α-平滑肌肌动蛋白。B—catenin胞质和核的异位表达阳性率为47．8％（33／69）。8号染色体三体阳性率为30．0％（6／20）,原发病例中8号染色体三体阳性率为1／12,显著低于复发病例的5／8。结论韧带样型纤维瘤病是主要发生于年轻女性的中间型肿瘤。它主要由纤维母细胞和肌纤维母细胞构成,而肌纤维母细胞有应力纤维、纤维连接复合体等特征性结构。可用荧光原位杂交方法在石蜡组织中检测8号染色体三体,其可能为韧带样型纤维瘤病的复发预测因子,并界定某种高复发风险亚型。     

乳腺癌HER2蛋白表达阳性者的基因状态分析

目的探索中国大陆女性乳腺癌HER2蛋白过表达人群中HER2基因状态、蛋白表达与基因扩增的一致性、17号染色体非整体性的发生情况及其意义。方法采用PathVysion^TM探针试剂盒,以荧光原位杂交（FISH）方法,分析经免疫组织化学（IHC）方法（HercepTest^TM试剂盒）确认的120例HER2蛋白表达阳性（IHC2＋者42例,IHC3＋者78例）的乳腺癌石蜡切片标本的HER2基因状态和17号染色体非整体性的发生率。结果HER2表达IHC2＋的标本中,基因扩增率76．19％（32／42）,其中低度扩增（比值2～4）26．19％（11／42）,中度扩增（比值4—10）41．62％（20／42）,高度扩增（比值〉10）2．38％（1／42）;IHC3＋的标本中,基因扩增率91．03％（71／78）,其中低度扩增11．54％（9／78）,中度扩增61．54％（48／78）,高度扩增17．95％（14／78）。全部病例中具有17号染色体非整体性者69．17％（83／120）,包括亚二体性（17号染色体平均数≤1．75）11．67％（14／120）、低多体性（17号染色体平均数2．26～3．75）43．33％（52／120）和高多体性（17号染色体平均数≥3.76）14．17％（17／120）。IHC2＋病例中17号染色体非整体性59．52％（25／42）,包括亚二体性7．14％（3／42）、低多体性42．86％（18／42）和高多体性9．52％（4／42）;IHC3＋病例中17号染色体非整体性74．36％（58／78）,包括亚二体性14．10％（11／78）、低多体性43．59％（34／78）和高多体性16．67％（13／78）。IHC2＋者FISH阳性以HER2基因中、低度扩增为主,而IHC3＋者则呈中、高度基因扩增的趋势,两者差异有统计学意义（P=0．0003）。IHC3＋中,7例FISH阴性中6例有17号染色体非整体性;IHC2＋中,10例FISH阴性中5例有17号染色体非整体性。结论在本组中国大陆女性乳腺癌人群中,IHC3＋与FISH阳性的一致性较高;IHC2＋者FISH阳性率高于国外研究者的数据;HER2蛋白阳性乳腺癌中17号染色体非整体性发生比例较高;IHC2＋和3＋者都具亚二体性、低多体性和高多体性特征,且两者均以低多体性为常见;IHC2＋者的HER2基因扩增多为中、低度扩增,而IHC3＋中FISH阳性者则以高比值者居多。IHC3＋而FISH阴性的主要原因为17号染色体非整体性。     

The ‘Morbid Anatomy’ of the Human Genome: Tracing the Observational and Representational Approaches of Postwar Genetics and Biomedicine The William Bynum Prize Essay

This paper explores evolving conceptions and depictions of the human genome among human and medical geneticists during the postwar period. Historians of science and medicine have shown significant interest in the use of informational approaches in postwar genetics, which treat the genome as an expansive digital data set composed of three billion DNA nucleotides. Since the 1950s, however, geneticists have largely interacted with the human genome at the microscopically visible level of chromosomes. Mindful of this, I examine the observational and representational approaches of postwar human and medical genetics. During the 1970s and 1980s, the genome increasingly came to be understood as, at once, a discrete part of the human anatomy and a standardised scientific object. This paper explores the role of influential medical geneticists in recasting the human genome as being a visible, tangible, and legible entity, which was highly relevant to traditional medical thinking and practice. I demonstrate how the human genome was established as an object amenable to laboratory and clinical research, and argue that the observational and representational approaches of postwar medical genetics reflect, more broadly, the interdisciplinary efforts underlying the development of contemporary biomedicine.     

The Art and Applications of Fluorescence In Situ Hybridization in Endocrine Pathology

Fluorescence in situ hybridization (FISH) or molecular cytogenetics is currently recognized as a reliable, sensitive, and reproducible technique for identifying the copy number and structure of chromosomes. FISH combines molecular genetics with classic cytogenetics and allows simultaneous morphologic evaluation on a single slide. Centromeric DNA probes are used to detect specific chromosomes and telomeric probes to demonstrate all chromosomes. Sequence-specific probes can localize in situ a single gene copy on a specific chromosome locus. FISH allows cytogenetic investigation of metaphase spreads and interphase nuclei. Several protocols have been proposed to analyze preparations from fresh samples or archival material. Comparative genomic hybridization (CGH) is a novel cytogenetic technique, which combines FISH with automatic digital image analysis. Comparative analysis of the hybridization products of tumor DNA and reference DNA with normal metaphase chromosomes, each labeled with color different fluorochrome, can retrieve chromosomal imbalances of the entire genome in a single experiment. FISH and CGH are powerful morphologic tools in understanding physiologic mechanisms and in resolving problems of the pathogenesis of several diseases. These techniques shed light on the cytogenetic background in many endocrinological disorders, providing a better understanding of the activities and alterations of endocrine cell function.     

Morphological,immunocytochemical and growth characteristics of three human glioblastomas established in vitro

Summary The human glioblastoma-derived cell lines 86HG-39, 87HG-28 and 87HG-31, used for the production of monoclonal antibodies (mAbs) against glioma-associated antigens (GAA), were characterized in terms of morphology, growth behaviour, chromosomes and antigen expression. In the primary tumours, differential expression of glial fibrillary acidic protein, S100 protein, Leu-7 and GAA as defined by mAbs MUC 2–39, MUC 2–63 and MUC 8–22 was demonstrated. Receptors for epidermal growth factor (EGFr) and nerve growth factor (NGFr) were found in many cells in short-term cultures, but the transferrin receptor (Tr) was found in only a few cells of 87HG-28. In permanent cell lines, differentiation antigens and EGFr decreased and Tr increased markedly. NGFr and GAA remained stable. Transplantation tumours of 86HG-39 were partly positive for Tr and GAA. Chromosomal analysis revealed that the 86HG-39 and 87HG-28 cell lines had a hypodiploid or diploid stem line with lines in the hypotetraploid to tetraploid region for 50 in vitro passages. The 87HG-31 cell line had chromosomal patterns in the hypotriploid to triploid region. A gain of chromosomes was seen in the groups C7, C8, C10, D14, F19, F20, G21, G22. The variability of antigens in these tumours and especially during long-term cultivation probably reveals an ability to influence the growth of malignant glioma cells via the respeective effector molecules.     

一例染色体复杂易位致胎儿多发畸形的遗传学诊断

目的: 对一例染色体复杂易位致多发畸形胎儿进行遗传学分析和诊断。方法: 对一例多发畸形胎儿行G显带染色体核型分析、单核苷酸多态性微阵列（SNP array）及荧光原位杂交（FISH）检测。胎儿父母行外周血染色体核型分析及FISH检测。结果: 胎儿的羊水染色体核型为46,XN,t（12;13）（q22;q32）。SNP array显示胎儿存在1q42.13q44重复（20 192 kb）及15q26.1q26.3缺失（13 293 kb）,核型分析与基因芯片结果不一致。FISH验证了SNP array的结果。母亲外周血FISH结果确认为隐匿性46,XX,t（1;15）（q42.1;q26.1）携带者,而胎儿遗传了其中一条衍生的15号染色体der（15）t（1;15）（q42.1;q26.1）。即胎儿遗传了父亲的t（12;13）（q22;q32）平衡易位及母亲的隐匿性平衡易位形成的衍生15号染色体。结论: 1q42.13q44重复和15q26.1q26.3缺失是导致本例胎儿畸形的遗传学病因,产前诊断时多种遗传学技术联合应用可为临床提供准确的诊断。     

八例22q11.2区微重复胎儿产前诊断和妊娠结局分析

目的: 探讨22q11.2区微重复与临床表型之间的关系,为遗传咨询提供依据。方法: 对2015年2月至2017年3月在杭州市妇产科医院、丽水市妇幼保健院产前诊断中心接受羊水穿刺产前诊断,染色体微阵列分析（CMA）报告为22q11.2区微重复的8例胎儿的产前诊断指征、胎儿超声检查情况、染色体核型、父母CMA检测结果、妊娠结局、出生后的生长发育情况进行回顾性分析。结果: 8例胎儿产前血清学筛查结果21三体高风险6例、胎儿颈项皮肤皱褶（NF）增厚1例、母亲染色体平衡易位1例。胎儿超声检查结果胎儿NF增厚1例,胎儿侧脑室增宽1例,未发现异常6例。CMA检测结果提示,22q11.2区域微重复片段大小为651 kb~3.2 Mb。6例胎儿通过父母的CMA溯源,均来自正常表型的父母一方,2例父母拒绝溯源。引产2例,继续妊娠6例。继续妊娠胎儿出生时外观正常,随访至今最大年龄3.5岁,生长发育和心理发育正常5例,生长迟缓1例。结论: 本组22q11.2区微重复胎儿出生前后无特异的临床表现,均遗传自无任何异常症状的父母一方,提示对22q11.2微重复胎儿应当慎重处理。     

四硫化四砷联合维甲酸、蒽环类药物治疗体系下附加染色体对急性早幼粒细胞白血病预后的影响

目的 分析在以维甲酸联合四硫化四砷、蒽环类药物为主的治疗体系下附加染色体对预后的影响以及有附加染色体异常患者的临床表现和对含四硫化四砷诱导治疗方案的反应性.方法 回顾性分析158例初治急性早幼粒细胞白血病患者临床资料.结果 附加染色体异常在急性早幼粒细胞白血病中占18.4%.其中三体8以及i17q-是最常见的附加异常,有附加染色体异常组患者弥散性血管内凝血发生率增高(P<0.05).在附加染色体异常组完全缓解率降低(75.9%vs 90.7%)、复发率增高(13.8%vs 6.2%)、中枢神经系统白血病的发生率增高(17.2%vs 6.2%),但差异未见统计学意义.两组生存率差异无统计学意义(P=0.160).结论 在此三类药物治疗体系下,尚未发现附加染色体对预后影响的统计学意义.     

染色体相互易位携带者的性别和易位片段长度对生育的影响

目的　探讨染色体相互易位携带者的性别和易位片段长度与生育的关系 ,为遗传咨询提供参考。　方法　收集相互易位核型 714例 ,综合分析涉及染色体不同部位的染色体异常和易位携带者性别对妊娠结局的影响。　结果　相互易位携带者妊娠结局以孕早期流产为主 ;随着易位片段增长 ,孕早期流产的比例增加 ,而生育染色体异常后代的风险减小 ;女性易位携带者较男性易位携带者易生育染色体异常后代 ,而男性易位携带者易出现不同程度不育。　结论　为易位携带者进行遗传咨询和生育风险估计时 ,要考虑易位的类型、易位片段的长度、已有的生育史和易位携带者的性别。