Silver STR Ⅲ系统基因座中等位基因在葡萄胎基因组中的分布及其意义

目的研究S ilver STR(Ⅲ系统3个基因座中各等位基因在葡萄胎基因组中的分布及潜在意义。方法对应用多重PCR和变性聚丙烯酰胺凝胶电泳技术鉴别出的39例遗传物质完全来自父方的完全性葡萄胎进行分析,统计S ilverSTR(Ⅲ系统3个基因座中各等位基因的出现率。结果在39例葡萄胎中,S ilver STR(Ⅲ系统的D16S539座位出现6个等位基因,D7S820座位出现5个等位基因,D13S317座位出现7个等位基因;D16S539座位中等位基因14出现率显著高于人群(P<0.05),D7S820座位中等位基因9出现率显著低于人群(P<0.01),其余各等位基因出现率与人群基因频率无显著差异;D16S539、D7S820和D13S317基因座位杂合度分别为0.1795,0.1282和0.1538;其中可追踪的23例良性葡萄胎中,D16S539出现6个等位基因,D7S820出现4个等位基因,D13S317出现7个等位基因;可追踪的7例侵袭性葡萄胎中,D7S820出现5个等位基因,D16S539与D13S317均出现4个等位基因,其中D16S539和D13S317座位的等位基因9出现率高于良性葡萄胎(P<0.01和P<0.05)。结论葡萄胎D16S539、D7S820和D13S317基因座位等位基因杂合度远低于人群杂合度;葡萄胎具有侵袭性与D16S539和D13S317座位的等位基因9出现率具有相关性。     

Silver STR® Ⅲ系统基因座中等位基因在葡萄胎基因组中的分布及其意义

目的研究Silver STR® Ⅲ系统3个基因座中各等位基因在葡萄胎基因组中的分布及潜在意义.方法对应用多重PCR和变性聚丙烯酰胺凝胶电泳技术鉴别出的39例遗传物质完全来自父方的完全性葡萄胎进行分析,统计Silver STR® Ⅲ系统3个基因座中各等位基因的出现率.结果在39例葡萄胎中,Silver STR® Ⅲ系统的D16S539座位出现6个等位基因,D7S820座位出现5个等位基因,D13S317座位出现7个等位基因;D16S539座位中等位基因14出现率显著高于人群(P＜0.05),D7S820座位中等位基因9出现率显著低于人群(P＜0.01),其余各等位基因出现率与人群基因频率无显著差异;D16S539、D7S820和D13S317基因座位杂合度分别为0.1795,0.1282和0.1538;其中可追踪的23例良性葡萄胎中,D16S539出现6个等位基因,D7S820出现4个等位基因,D13S317出现7个等位基因;可追踪的7例侵袭性葡萄胎中,D7S820出现5个等位基因,D16S539与D13S317均出现4个等位基因,其中D16S539和D13S317座位的等位基因9出现率高于良性葡萄胎(P＜0.01和P＜0.05).结论葡萄胎D16S539、D7S820和D13S317基因座位等位基因杂合度远低于人群杂合度;葡萄胎具有侵袭性与D16S539和D13S317座位的等位基因9出现率具有相关性.     

Silver STR(R) Ⅲ系统基因座中等位基因在葡萄胎基因组中的分布及其意义

目的研究Silver STR(R) Ⅲ系统3个基因座中各等位基因在葡萄胎基因组中的分布及潜在意义.方法对应用多重PCR和变性聚丙烯酰胺凝胶电泳技术鉴别出的39例遗传物质完全来自父方的完全性葡萄胎进行分析,统计Silver STR(R) Ⅲ系统3个基因座中各等位基因的出现率.结果在39例葡萄胎中,Silver STR(R) Ⅲ系统的D16S539座位出现6个等位基因,D7S820座位出现5个等位基因,D13S317座位出现7个等位基因;D16S539座位中等位基因14出现率显著高于人群(P＜0.05),D7S820座位中等位基因9出现率显著低于人群(P＜0.01),其余各等位基因出现率与人群基因频率无显著差异;D16S539、D7S820和D13S317基因座位杂合度分别为0.1795,0.1282和0.1538;其中可追踪的23例良性葡萄胎中,D16S539出现6个等位基因,D7S820出现4个等位基因,D13S317出现7个等位基因;可追踪的7例侵袭性葡萄胎中,D7S820出现5个等位基因,D16S539与D13S317均出现4个等位基因,其中D16S539和D13S317座位的等位基因9出现率高于良性葡萄胎(P＜0.01和P＜0.05).结论葡萄胎D16S539、D7S820和D13S317基因座位等位基因杂合度远低于人群杂合度;葡萄胎具有侵袭性与D16S539和D13S317座位的等位基因9出现率具有相关性.     

P57和Ki-67在葡萄胎鉴别诊断中的作用

目的:探讨P57和Ki-67蛋白在完全性和部分性葡萄胎鉴别诊断中的作用.方法:分别收集正常胎盘绒毛、部分性葡萄胎和完全性葡萄胎石蜡标本各12例,应用免疫组织化学方法检测P57和Ki-67蛋白在这些病变中的分布及表达水平.结果:P57蛋白在正常绒毛及部分性葡萄胎组织中主要分布于绒毛的细胞滋养叶细胞及间质细胞,两组间阳性率...     

葡萄胎中c-myc和ras癌基因表达及其意义

目的　研究探讨葡萄胎中c myc、ras基因表达情况及其意义。方法　采用免疫组织化学染色对 4 2例葡萄胎和10例早孕绒毛标本中c myc、ras基因表达进行检测 ,并采用图像分析技术 ,对正常早孕绒毛组和葡萄胎不同病理分级组间c myc、ras的表达情况进行对比分析。结果　c myc、ras阳性信号表达在正常早孕绒毛和葡萄胎Ⅰ Ⅲ级各组间均无明显统计学差异 (P >0 0 5 )。结论　葡萄胎中c myc、ras癌基因表达情况与病理学分级无关 ,与正常绒毛也无统计学差异 ,支持以下观点 :即大多数葡萄胎只是流产的一种形式而非真正的肿瘤。     

p57Kip2在完全性和部分性葡萄胎与流产伴绒毛水肿中的表达及意义

p57^Kip2是细胞周期抑制因子和肿瘤抑制基因,定位于染色体11p15．5。其所编码产物是一种广泛的细胞周期蛋白依赖性激酶（CDK）的抑制蛋白。p57^Kip2显示极强的父本基因组烙印,在母本等位基因中为优势,即该基因的表达是由母系来源的基因开放所致。我们利用免疫组织化学法检测p57^Kip2蛋白在由组织学所诊断的完全性葡萄胎、部分性葡萄胎及流产伴绒毛水肿中的表达情况,试图探讨p57^Kip2基因在完全性、部分性葡萄胎和流产伴绒毛水肿鉴别诊断中的临床意义。     

nm23蛋白在乳腺癌转移和葡萄胎恶变中的研究

目的:探讨nm23蛋白在乳腺癌及乳腺癌转移;葡萄胎与葡萄胎恶性变中的关系。方法:应用斑点ELISA法,检测81例临床、病理确诊为乳腺癌病人和18例葡萄胎(其中8例临床,病理确诊为恶性葡萄胎)。病人nm23基因表达水平。结果:81例乳腺癌中53例阳性表达(65.4%),28例为阴性表达(34.5%)。而阴性表达的28例中有19例临床、病理和X线证实为乳腺腺癌转移,两者符合率为67.8%(19/28),18例葡萄胎患者中,临床、病理已证实为恶性葡萄胎的8例患者,其nm23蛋白呈减低表达的有3例,完全阴性表达的3例,而阳性表达的仅有2例,符合率75%(6/8)。而未发现恶性变的10例葡萄胎患者仅有1例显示弱阳性表达,其余9例均呈阳性表达,两者差别显著(P<0.001)。结论:斑点ELISA法由于方法简便、快速、易于取材,可作为临床早期判断肿瘤预后的一个重要生化检测指标。     

PCNA与Ki-67在葡萄胎患者宫内不同部位组织中的表达及临床意义

目的探讨PCNA和Ki-67在葡萄胎患者宫内不同部位组织中的表达对葡萄胎治疗及预后判断的意义.方法采用免疫组化S-P法检测38例葡萄胎患者宫腔中央水泡样胎块及靠宫壁蜕膜样组织中PCNA和Ki-67的表达.结果 1.PCNA及Ki-67在宫中央组织阳性表达均大于靠宫壁组织.P＜0.05,有明显差异.2.完全性葡萄胎PCNA及Ki-67阳性表达>部分性葡萄胎.浸润性葡萄胎PCNA及Ki-67阳性表达>完全性葡萄胎,且均为强阳性表达.3.PCNA和Ki-67在两组织中均有阳性表达,PCNA的表达更为明显,P＜0.05(宫内)P＜0.05(宫壁),有显著差异.结论葡萄胎宫内水泡样组织的增殖活性明显大于靠宫壁的组织,必须彻底刮净宫内组织方能预防恶变的发生.靠宫壁组织因其增殖活性较弱,可不必常规二次刮宫,以避免过度搔刮带来的过度损伤.葡萄胎的细胞增殖程度可能随疾病进展而逐渐增强,对于宫内、宫壁组织均有强阳性表达的患者要警惕其恶变的可能.PCNA与Ki-67能代表葡萄胎滋养细胞异常增生的客观指标,其检测简便、快捷、价廉,有临床价值,值得推广.     

P57 kip2在葡萄胎中表达及临床意义

目的探讨P57 kip2在完全性葡萄胎(CM)、部分性葡萄胎(PM)及水肿流产绒毛(HA)诊断中的作用。方法应用免疫组化的方法检测P57蛋白在完全性葡萄胎28例,部分性葡萄胎30例,水肿绒毛30例组织中的表达,并应用胶体金免疫电镜技术观察P57 kip2在以上组织中的超微结构定位。结果 P57 kip2在30例HA 27例(90%)呈阳性表达,30例PM 25例(83.33%)呈阳性表达,28例CM 3例(10.71%)呈阳性表达,并且在CM中的阳性强度均较弱。3种病变P57 kip2染色阳性率统计分析显示CM与PM及HA之间均存在显著性差异(P<0.01),而PM与HA之间无显著性差异(P>0.05)。完全性葡萄胎细胞内质网和核膜上有黑色圆形集中的胶体金颗粒分布,部分性葡萄胎病变组织中无发现胶体金颗粒分布。结论 P57 kip2免疫组化是诊断CM的一种高敏感性、高特异性方法,同时用胶体金免疫电镜技术检测P-57kip2蛋白在CM的超微结构定位,对早期CM的发现有重大作用。     

Single haplotype assembly of the human genome from a hydatidiform mole

A complete reference assembly is essential for accurately interpreting individual genomes and associating variation with phenotypes. While the current human reference genome sequence is of very high quality, gaps and misassemblies remain due to biological and technical complexities. Large repetitive sequences and complex allelic diversity are the two main drivers of assembly error. Although increasing the length of sequence reads and library fragments can improve assembly, even the longest available reads do not resolve all regions. In order to overcome the issue of allelic diversity, we used genomic DNA from an essentially haploid hydatidiform mole, CHM1. We utilized several resources from this DNA including a set of end-sequenced and indexed BAC clones and 100× Illumina whole-genome shotgun (WGS) sequence coverage. We used the WGS sequence and the GRCh37 reference assembly to create an assembly of the CHM1 genome. We subsequently incorporated 382 finished BAC clone sequences to generate a draft assembly, CHM1_1.1 (NCBI AssemblyDB GCA_000306695.2). Analysis of gene, repetitive element, and segmental duplication content show this assembly to be of excellent quality and contiguity. However, comparison to assembly-independent resources, such as BAC clone end sequences and PacBio long reads, indicate misassembled regions. Most of these regions are enriched for structural variation and segmental duplication, and can be resolved in the future. This publicly available assembly will be integrated into the Genome Reference Consortium curation framework for further improvement, with the ultimate goal being a completely finished gap-free assembly.The production of a reference sequence assembly for the human genome was a milestone in biology and clearly has impacted many areas of biomedical research (McPherson et al. 2001; International Human Genome Sequencing 2004). The availability of this resource allows us to investigate genomic structure and variation at a depth previously unavailable (Kidd et al. 2008; The 1000 Genomes Project Consortium 2012). These studies have helped make clear the shortcomings of our initial assembly models and the difficulty of comprehensive genome analysis. While the current human reference assembly is of extremely high quality and is still the benchmark by which all other human assemblies must be compared, it is far from perfect. Technical and biological complexity lead to both missing sequences as well as misassembled sequence in the current reference, GRCh38 (Robledo et al. 2002; Eichler et al. 2004; International Human Genome Sequencing 2004; Church et al. 2011; Genovese et al. 2013).The two most vexing biological problems affecting assembly are (1) complex genomic architecture seen in large regions with highly homologous duplicated sequences and (2) excess allelic diversity (Bailey et al. 2001; Mills et al. 2006; Korbel et al. 2007; Kidd et al. 2008; Zody et al. 2008). Assembling these regions is further complicated due to the fact that regions of segmental duplication (SD) are often correlated with copy-number variants (CNVs) (Sharp et al. 2005). Regions harboring large CNV SDs have been misrepresented in the reference assembly because assembly algorithms aim to produce a haploid consensus. Highly identical paralogous and structurally polymorphic regions frequently lead to nonallelic sequences being collapsed into a single contig or allelic sequences being improperly represented as duplicates. Because of this complexity, a single, haploid reference is insufficient to fully represent human diversity (Church et al. 2011).The availability of at least one accurate allelic representation at loci with complex genomic architecture facilitates the understanding of the genomic architecture and diversity in these regions (Watson et al. 2013). To enable the assembly of these regions, we have developed a suite of resources from CHM1, a DNA source containing a single human haplotype (Taillon-Miller et al. 1997; Fan et al. 2002). A complete hydatidiform mole (CHM) is an abnormal product of conception in which there is a very early fetal demise and overgrowth of the placental tissue. Most CHMs are androgenetic and contain only paternally derived autosomes and sex chromosomes resulting either from dispermy or duplication of a single sperm genome. The phenotype is thought to be a result of abnormal parental contribution leading to aberrant genomic imprinting (Hoffner and Surti 2012). The absence of allelic variation in monospermic CHM makes it an ideal candidate for producing a single haplotype representation of the human genome. There are a number of existing resources associated with the “CHM1” sample, including a BAC library with end sequences generated with Sanger sequencing using ABI 3730 technology (https://bacpac.chori.org/), an optical map (Teague et al. 2010), and a BioNano genomic map (see Data access), some of which have previously been used to improve regions of the reference human genome assembly.BAC clones have historically been used to resolve difficult genomic regions and identify structural variants (Barbouti et al. 2004; Carvalho and Lupski 2008). A BAC library constructed from CHM1 DNA (CHORI-17, CH17) has also been utilized to resolve several very difficult genomic regions, including human-specific duplications at the SRGAP2 gene family on Chromosome 1 (Dennis et al. 2012). Additionally, the CHM1 BAC clones were used to generate single haplotype assemblies of regions that were previously misrepresented because of haplotype mixing (Watson et al. 2013). Both of these efforts contributed to the improvement of the GRCh38 reference human genome assembly, adding hundreds of kilobases of sequence missing in GRCh37, in addition to providing an accurate single haplotype representation of complex genome regions.Because of the previously established utility of sequence data derived from the CHM1 resource, we wished to develop a complete assembly of a single human haplotype. To this end, we produced a short read-based (Illumina) reference-guided assembly of CHM1 with integrated high-quality finished fully sequenced BAC clones to further improve the assembly. This assembly has been annotated using the NCBI annotation process and has been aligned to other human assemblies in GenBank, including both GRCh37 and GRCh38. Here we present evidence that the CHM1 genome assembly is a high-quality draft with respect to gene and repetitive element content as well as by comparison to other individual genome assemblies. We will also discuss current plans for developing a fully finished genome assembly based on this resource.     

Serum protein distribution in hydatidiform mole. An immunohistochemical study

In normal placentas during the first trimester of pregnancy, the syncytiotrophoblast appeared to be immunoreactive to alpha-antitrypsin (alpha 1-AT), alpha 1-antichymotrypsin, albumin, IgG, and transferrin. The underlying cytotrophoblast was negative for these same serum proteins. In the hydatidiform mole, these findings were profoundly different. The syncytiotrophoblast lost its immunoreactivity to albumin, IgG, transferrin, and, less frequently, to alpha 1-AT. Furthermore, the underlying cytotrophoblast became immunoreactive to albumin, alpha 1-AT, IgG, transferrin, ferritin, orosomucoid, and, sometimes, to alpha 1-antichymotrypsin. This altered immunohistochemical pattern suggested a notable change in the pinocytotic activity of the trophoblast in the placenta during molar degeneration. The absence of pinocytosis in the syncytiotrophoblast for several proteins can be explained by the partial loss of specific membrane receptors. The contemporaneous appearance of numerous serum proteins in the cytotrophoblast could indicate an activation, not only proliferative, but also functional, in the germinative cytotrophoblast. Diagnostically, this histochemical finding in the hydatidiform mole, which was quite different from that seen in normal placentas during the first trimester of pregnancy, could provide additional evidence concerning trophoblastic abnormalities in the chorionic villi during molar degeneration.     

Distribution of the alleles at loci D16S539, D7S820, and D13S317 in hydatidiform mole genome from Chinese women and its relationship with clinical prognosis

Using polymerase chain reaction and denaturating polyacrylamide gel electrophoretic techniques, we studied 53 cases of hydatidiform moles. Of these, 41 cases were genetically complete hydatidiform moles (g-CHM) whose genome were totally paternally derived. We investigated the distribution of the alleles in the short tandem repeat sequences at loci D16S539, D7S820, and D13S317 in these cases. In particular, we analyzed the allelic distribution and potential significance in cases with traceable benign and invasive moles (i.e., persistent trophoblastic tumor [PTT]). Among 41 g-CHM cases, there were six alleles at D16S539, five alleles at D7S820 (the frequencies of alleles 9 and 10 were respectively lower and higher than those in Beijing population), and seven alleles at D13S317; the heterozygosity of loci D16S539, D7S820, and D13S317 was 0.0732, 0.0976, and 0.0732, respectively. Among 23 benign cases, there were six alleles at D16S539, four at D7S820, and six at D13S317; among 11 PTT cases, there were five alleles at D7S820 and four alleles each at D16S539 and D13S317. The frequencies of allele 9 at D16S539 and allele 10 at D7S820 were higher than in benign cases (P < 0.05). There were significant differences in frequencies of alleles 9 and 10 at D7S820 between the cases and the Beijing population, and heterozygosity at the three loci was lower in the cases than in the population. In addition, invasiveness of hydatidiform mole correlated to the frequency of allele 9 at loci D16S539 and allele 10 at D7S820.     

葡萄胎绒毛组织VEGF及bFGF表达与血管缺失

目的：探讨血管内皮生长因子（VEGF）及碱性成纤维细胞生长因子（bFGF）与葡萄胎血管缺失的关系。 方法: 采用免疫组化S-P法检测正常早孕绒毛及葡萄胎绒毛中CD34、VEGF与bFGF的表达强度并观察葡萄胎绒毛间质血管形态学特征。 结果: （1）完全性葡萄胎绒毛间质仍可见为数不少的血管。（2）完全性葡萄胎组微血管密度（MVD）明显低于部分性葡萄胎组(P<0.01)，部分性葡萄胎组MVD明显低于正常早孕组(P<0.01)。（3）绒毛间质，完全性葡萄胎组VEGF、bFGF表达明显低于部分性葡萄胎组(均P<0.01)，部分性葡萄胎组VEGF、bFGF表达明显低于正常早孕组(均P<0.01)。 结论: 葡萄胎绒毛间质VEGF及bFGF低表达与血管缺失有关。葡萄胎血管发育障碍，导致胚胎死亡。