应用荧光原位杂交技术快速诊断胎儿染色体数目异常

目的 探讨荧光原位杂交（ｆｌｕｏｒｅｃｅｎｔ ｉｎ ｓｉｔｕ ｈｙｂｒｉｄｉｚａｔｉｏｎ,ＦＩＳＨ）技术在快速产前诊断胎儿染色体数目异常中的价值。方法 对２０例孕１６￣３６周,有产前诊断指征者,在Ｂ超引导下经腹抽取羊水后,应用Ｘ、Ｙ、１８号染色体着丝粒探针１３ｑ１４－ｑ２１和２１ｑ１１特异性探讨,对未培养的羊水间期细胞进行ＦＩＳＨ,然后用荧光显微镜进行观察,并用Ａｐｐｌｉｅｄ ｉｍａｇｉｎｇ染色体     

性畸形四例遗传学分析

在细胞遗传学核型分析的基础上，利用Ｙ染色性体决定区基因（ｓｅｘ－ｄｅｔｅｒｍｉｎｉｇｒｅｇｉｏｎｏｆｔｈｅＹｃｈｒｏｍｏｓｏｍｅ，ＳＲＹ）探针，对４例性畸形病例进行了Ｓｏｕｔｈｅｒｎ杂交分析。同时运用ＳＲＹ编码区的特异性引物，对其基因组ＤＮＡ进行了聚合酶链式反应（ＰＣＲ）扩增检测。结果显示：１例４６，ＸＹ女性伴性腺发育不良症；１例４５，ＸＯ／４６，ＸＹ女性表现为Ｔｕｒｎｅｒ综合征，未检出ＳＲＹ特异     

多囊卵巢综合征患者血清生长激素和泌乳素对左旋多巴反应的研究

目的：探讨多囊卵巢综合征（ＰＣＯＳ）患者生长激素（ＧＨ）和泌乳素（ＰＲＬ）异常分泌的机理。方法：对黄体生成素（ＬＨ）／卵泡刺激素（ＦＳＨ）≥３的１５例患者（Ｉ型组）、ＬＨ／ＦＳＨ＜３的１５例患者（Ⅱ型组）以及２０例月经周期正常妇女（对照组），行左旋多巴（Ｌ－ＤＡ，５００ｍｇ）兴奋下丘脑－垂体轴功能试验，观察３组ＧＨ和ＰＲＬ的浓度变化。结果：在基础状态下，Ⅰ、Ⅱ型组的ＧＨ较低（Ｐ＜０．０１）、ＰＲＬ较高（Ｉ型组，Ｐ＜０．０５）；Ｌ－ＤＡ兴奋试验后，Ⅰ、Ⅱ型组ＧＨ的升高程度和ＰＲＬ的下降程度均低于对照组。结论：ＰＣＯＳ的ＧＨ和ＰＲＬ异常分泌可能与其中枢ＤＡ活性不足有关。     

体外受精-胚胎移植治疗不孕症85例报道

目的：评价应用体外受精－胚胎移植（ＩＶＦ－ＥＴ）治疗８５例不孕患者的临床结果。方法：用药方案有：①ＧｎＲＨａ／ＦＳＨ／ｈＭＧ；②ＧｎＲＨａ／ＦＳＨ；③ＧｎＲＨａ／ｈＭＧ。培养液包括Ｅａｒｌｅ’ｓ平衡盐液及人类输卵管液（ＨＴＦ）。结果：８５例进行了１０９个ＩＶＦ周期治疗，１２个周期（１１０％）因反应不良而取消。每周期获取卵母细胞（１０２±５４）个，卵子受精率６８９％，卵裂率６０８％，每个转移胚胎植入率７７％（３１／４０２）。９４个移植周期中，每周期移植（４９±２４）个胚胎，获２０次临床妊娠（２１３％）。采用ＨＴＦ妊娠率为２２４％，高于应用Ｅａｒｌｅ’ｓ液的妊娠率（１８５％），但差异无统计学意义。２０次妊娠患者中，１次输卵管妊娠，４次自然流产（２０％），５例足月分娩，１０例继续妊娠。多胎妊娠率４０％，重度ＯＨＳＳ发生率４１％。结论：ＩＶＦ－ＥＴ是治疗输卵管因素、子宫内膜异位症等不孕症的重要而有效的手段。体外培养时用ＨＴＦ可能优于Ｅａｒｌｅ’ｓ液，而有利于提高妊娠率。     

应用荧光原位杂交技术对罗伯逊易位携带者进行胚胎植入前诊断

罗伯逊易位是常见的染色体结构异常 ,患者常常表现为反复自然流产。胚胎植入前遗传学诊断 (ｐｒｅｉｍｐｌａｎｔａｔｉｏｎｇｅｎｅｔｉｃｄｉａｇｎｏｓｉｓ,ＰＧＤ) ,可以选择正常的胚胎进行移植 ,有利于获得正常妊娠从而达到优生优育的目的。我们应用荧光原位杂交 (ｆｌｕｏｒｅｓｃｅｎｃｅｉｎ ｓｉｔｕｈｙｂｒｉｄｉｚａｔｉｏｎ ,ＦＩＳＨ)方法对罗伯逊易位携带者进行ＰＧＤ ,现报道如下。一、资料与方法1.病例介绍 :患者 30岁 ,已婚 6年 ,反复自然流产 5次 ,外周血染色体核型分析为罗伯逊易位携带者 ,即 45 ,ＸＸ ,- 13,- 14, ｔ(…     

简并寡核苷酸引物聚合酶链反应扩增单细胞全基因组均匀性 …

目的 搪塞单细胞简交核苷酸经物聚合酶链反应（ＤＯＰ－ＰＣＲ）扩境界量基因组ＤＮＡ的均匀性及植入前胚胎遗传学研究的新途径。方法 获取正常男女性,Ｘ单体、Ｘ、１３和２１三体行单细胞ＤＯＰ－ＰＣＲ,通过比较基因组杂交（ＣＧＨ）双色荧光强度比变化,分析扩增均匀性。结果 （１）ＤＯＰ＝ＰＣＲ产物／正常男性组ＤＮＡＣＧＨ：约１／３常染色持区强度比均数及标准差超过正常允许范围,Ｘ染色体假性过度表达,１３、２１三     

三种助孕技术治疗多囊卵巢综合征不孕的疗效比较

目的：观察３种助孕技术：ＩＶＦ ＥＴ、ＧＩＦＴ和控制性超促排卵（ＣＯＨ）后自行同房对多囊卵巢综合征（ＰＣＯＳ）不孕患者的疗效。方法：应用３种方法治疗,观察各组的临床妊娠率和发生卵巢过度刺激综合征（ＯＨＳＳ）的情况。结果：ＩＶＦ ＥＴ和ＧＩＦＴ与ＣＯＨ相比能得到更高的临床妊娠率,而前二者发生ＯＨＳＳ的危险性明显低于后者。结论：为避免ＣＯＨ后发生ＯＨＳＳ,建议对因ＰＣＯＳ不孕的患者采取体外助孕技术治疗。应用何种技术要根据患者的具体情况而定。如果有至少１ 条输卵管完全正常,可先试行ＧＩＦＴ。此外,须将多余的卵子进行体外授精试验,观察其受精及卵裂情况,从而对今后的治疗提出指导性建议,并要将得到的胚胎进行冷冻保存,以备今后使用。     

Stuuyon thesecondrycytoreductivesurgeryforptientswithdvncedovrinepithelilcncer

１ＩＭＰＡＣＴＯＦＰＲＩＭＡＲＹＣＹＴＯＲＥＤＵＣＴＩＶＥＳＵＲＧＥＲＹＯｖɑｒɑｎｅｐｉｔｈｅｌｉɑｌｃɑｎｃｅｒ（ＯＥＣ）ｉｓｔｈｅｍｏｓｔｌｅｔｈɑｌｇｙｎｅｃｏｌｏｇｉｃｎｅｏｐｌɑｓｍ,ｌɑｒｇｅｌｙｄｕｅｔｏｆｒｅｑｕｅｎｔｐｒｅｓｅｎｃ...     

羟甲基无环鸟苷对携有单纯疱疹病毒胸腺嘧啶核苷激…

目的：观察羟甲基无环鸟苷（ＧＣＶ）对携有Ｉ型单纯疱疹病毒胸腺嘧淀核苷激酶基因（ＨＳＶ１－ｔｋ）的人卵巢上皮癌ＡＯ细胞的体内杀伤效应，方法；于裸级部左右两侧皮下分别接种ＡＯ细胞及携有ＨＳＶ１－ｔｋ基因的ＡＯ细胞（ＡＯ／ＨＳＶ１－ｔｋｃ），成瘤后每日腹腔注射ＧＣＶ。结果：经ＧＶＣ治疗后，裸鼠背部两侧肿瘤平均重量分别为：左侧ＡＯ肿瘤０．６８１ｇ；右侧ＡＯ／ＨＳＶ１－ｔｋｃ肿瘤０．０８７ｇ。经ＧＣＶ作用后     

胎儿胰岛素样生长因子-Ⅰ的检测及意义

目的评估胰岛素样生长因子Ⅰ（ＩＧＦⅠ）在胎儿、胎盘生长发育中的作用。方法用放射免疫分析法（ＲＩＡ）测定大于胎龄儿（ＬＧＡ）组３０例,适于胎龄儿（ＡＧＡ）组３６例及小于胎龄儿（ＳＧＡ）组３６例的脐血清ＩＧＦⅠ的水平。用线性相关分析法分析各组变量之间的相关关系。结果３组胎儿脐血清ＩＧＦⅠ与胎龄、胎儿体重、胎盘重量均呈显著正相关（ｒ＝０．３２,Ｐ＜０．００１;ｒ＝０．６８,Ｐ＜０．００１;ｒ＝０．７５,Ｐ＜０．０１）,其中与胎盘重量呈高度正相关。脐血清ＩＧＦⅠ水平,ＡＧＡ组为１４４５９±４６．７３μｇ／Ｌ;ＳＧＡ组为９０．８０μｇ／Ｌ（ｔ＝４．７,Ｐ＜０．００１）,ＬＧＡ组为２０９１７μｇ／Ｌ（ｔ＝７．９７,Ｐ＜０．００１）。结论ＩＧＦⅠ是胎儿、胎盘生长发育的重要调节因子,对巨大儿、小于胎龄儿的形成有重要作用。     

A familial Xp+ chromosome detected during fetal karyotyping,which is associated with short stature in four generations of a Turkish family

The short-stature homeobox-containing gene (SHOX) on chromosome Xp22.3 was recently identified as an important determinant of the stature phenotype. Deletions of the SHOX gene, some of them due to structural chromosome abnormalities, have been described in patients with idiopathic short stature and Leri-Weill syndrome. Additionally, haploinsufficiency of SHOX is a main cause for short stature seen in patients with Turner syndrome.Here we report an unusual X-chromosome abnormality, which was detected during a fetal karyotyping performed because of a previous child with Down syndrome. GTG banding demonstrated an extra chromosome segment on the terminal part of the short arm of chromosome X in the index case (karyotype: 46,X,Xp+). The same chromosomal abnormality was found in the mother and the maternal grandmother. All carriers of this chromosomal abnormality presented with short stature but no other associated symptoms.Whole chromosome painting of X revealed a homogeneous painting of the abnormal X chromosome indicating that no other chromosome was involved. Additional FISH studies with probe DXS1140 (Kallmann probe at Xp22.3), Quint-Essential X-Specific DNA (DMD probe at Xp21.2), XIST (at Xq13.2), and Tel Xq/Yq were performed, and no abnormality was observed in the intensities or the localizations of the probes signals. However, applying a specific SHOX gene probe (derived from cosmid LLNONO3M34F5) showed a loss of signal on the derivative X chromosome. Our results show that the Xp+ generation led to a deletion of the complete SHOX gene and caused short stature in the presented family.     

应用荧光原位杂交技术诊断羊水细胞染色体异常

探讨荧光源位杂交技术于产前诊断羊水细胞染色体异常的实验方法的应用价值。方法对３４例孕１６－２３周有产前认旨征者经腹部抽取羊水,用Ｃｈａｎｇ培养液传代培养,常规制备羊水细胞分裂中期染色体,应用生物素及地高辛标记的人类全着线粒探针和Ｘ、Ｙ、１３、２１、１８号染色体     

45,XO/46,X,dic(Yq) mosaicism in Turner's phenotype with endodermal sinus tumor of the ovary

A case of a endodermal sinus tumor of the ovary in a patient with 45,XO/46,X,dic(Y) mosaicism is reported because of the rarity of the karyotype and condition. A 15-year-old girl was admitted to Yamagata University Hospital because of constipation for several days. Physical examination showed webbing of the neck, cubitus valgus and short stature. Her abdomen was bulging. Chromosomal analysis showed 45,XO/46,X,dic(Yq) mosaicism in karyotype. alpha-Fetoprotein and CA-125 in the serum were high. A left ovarian tumor was found by laparotomy; however, the right ovary was a streak gonad and the uterus was hypoplastic. An endodermal sinus tumor was diagnosed by a pathologist. After operation, cisplatin-vinblastin-bleomycin chemotherapy was instituted and the tumor marker went down. This patient is still healthy and under observation at the outpatient clinic.     

1例嵌合型45,X/46,X,r(Y)患者的遗传学分析

目的:应用细胞遗传学和分子生物学技术分析1例嵌合型45,X/46,X,r(Y)患者的核型。方法:应用常规染色体标本制备方法进行G-显带和C-显带;并应用CEPX(DXZ1,Xp11.1-q11.1,Spectrum Green,Vysis)探针、LSI SRY(Yp11.3,Spectrum Orange,Vysis)探针和CEP18(D18Z1,18p11.1-q11.1,Spectrum Aqua,Vysis)与患者的中期分裂相进行荧光原位杂交(fluorescence in situ hybridization,FISH);同时应用PCR技术对患者进行Y染色体微缺失检测。结果:结合G-显带、C-显带、FISH检测结果和Y染色体微缺失的检测结果,确定该患者核型为46,X,r(Y)(p11.3q12)[85]/45,X[15]。Yq11区生精基因微缺失检测未显示该患者存在缺失。结论:细胞遗传学检测结合FISH可以诊断复杂的染色体异常,为患者提供正确的遗传咨询和生育指导。     

Molecular and cytogenetic characterization of extra-structurally abnormal chromosomes (ESACs) found prenatally: outcome and follow-up

A 40-year-old woman underwent amniocentesis at 15.3 weeks of gestation. Chromosome analysis performed using QFQ, DA-DAPI and CBG banding revealed two de novo extra-chromosomal markers (ESACs) in 11 of the 16 colonies analysed. Fluorescence in situ hybridization (FISH) showed that both chromosomes came from the Yq11.22.1 region of the Y chromosome. PCR analysis of genes and STS localized on the Y chromosome excluded the Yp presence specifically of the SRY gene, and most of the euchromatic region of Yq. After extensive genetic counselling and considering both laboratory and second-level ultrasound data, the couple decided to continue the pregnancy. At 37.4 weeks of gestational age, a girl weighing 2750 g was born with an Apgar score of 9/10. A blood sample taken from the umbilical cord showed three cellular lines: mos47,XX, +mar1 ish.der (Y)(wcpY+) [21%]/48,XX, +mar1 ish.der (Y)(wcpY+), +mar2 ish.der (Y)(wcpY+) [41%]/46,XX [38%]. One year after birth, the baby was developing normally and had normal psychomotorial activity.     

Turner综合征患者标记染色体的鉴定与分析

目的 分析11例携带标记染色体的Turner综合征患者的核型，研究这类染色体的表型效应。方法 选择11例具Turner综合征表型的患者，常规核型分析均显示为携带标记染色体的嵌合体，其中6例标记染色体呈环状。患者G带核型表示为mos．45，X／46，X，＋mar或者mos．45，X／46，X，＋r．以X／Y着丝粒探针，应用荧光原位杂交（FISH）技术分析这些标记染色体起源，对其中2例较大的环状染色体，结合反向染色体涂染确定断裂位点，比较不同断裂位点的标记染色体的遗传学效应。结果11例患者所携带的标记染色体均为环状染色体，r（X）的断裂位点分别位于Xp22、Xq22、Xq24、Xq26等。结论 Turner综合征患者的标记染色体主要来源于X染色体，且表现为r（X）形式。r（X）均以嵌合型的形式存在。     

45,X/46,X,del(Yq) sex chromosome mosaicism--analysis of the phenotypic expression

Three female patients with Turner-syndrome (sexual infantilism, short stature and somatic Turner-stigmata) have been analysed cytogenetically by means of different banding techniques. A deletion of the distal heterochromatic band Yq12 of the Y chromosome was observed in a mosaic with a 45,X-cell line, i.e. the karyotype is 45,X/46,X,del(Y)(q12). In order to get information about the phenotypic expression of the 45,X/46,X,del(Yq) mosaicism all previously published cases have been reviewed. Comparing the phenotypes of all 45,X/46,X,del(Yq) mosaic cases three different phenotype categories of sexual development can be distinguished: female individuals with sexual infantilism and Turner-stigmata, individuals with ambiguous genitals, ranging from clitoris hypertrophy of female genitals to hypospadia of males, male individuals, who are infertile (azoospermic). A comparison of the appearance of external genitals with the status of gonads of all patients revealed an unequivocal relationship between the gonad status and the resulting phenotype category. Furthermore, the role of Y-chromosomal loci determining testicular differentiation (biological function of H-Y antigen) for male development has been emphasized. The effect of the 45,X-cell line on the expression of short stature and somatic Turner-stigmata is independent of sexual development. Considering the great phenotypic variability of the 45,X/46,X,del(Yq) mosaicism it seems impossible to deduce a definitive phenotype. This problem is acute in prenatal diagnosis especially.     

Y染色体长臂缺失及不分离不育男性1例报道

目的:报道1例Y染色体长臂缺失合并不分离的男性无精子症患者。方法:常规染色体核型分析,荧光原位杂交以确定核型。PCR-STSs检测以确定Y染色体断裂点,并行睾丸活检。结果:细胞遗传学和FISH证实患者为嵌合体,核型为45,X/46,X,del(Y)/47,X,del(Y)del(Y)。分别占27%,68%,5%。C带显示患者Yq12全部丢失。PCR-STSs检测AZFa存在,AZFb和AZFc区域全部丢失,断裂点位于sY88和sY95之间及sY88以下。睾丸病理显示精曲小管中只有支持细胞,没有生精细胞。未见卵巢组织。结论:患者无精子症、睾丸体积小与病理结果一致,其原因是由于Yq11.2的缺失。     

Prenatal and postnatal characterization of Y chromosome structural anomalies by molecular cytogenetic analysis

We describe three cases in which we used fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR) and comparative genomic hybridization (CGH) to characterize Y chromosome structural anomalies, unidentifiable by conventional G-banding. Case 1 was a 46,X,+mar karyotype; FISH analysis revealed an entire marker chromosome highlighted after hybridization with the Y chromosome painting probe. The PCR study showed the presence of Y chromosome markers AMG and SY620 and the absence of SY143, SY254 and SY147. CGH results confirmed the loss of Yq11.2-qter. These results indicated the presence of a deletion: del(Y)(q11.2). Case 2 was a 45,X [14]/46,XY[86] karyotype with a very small Y chromosome. The PCR study showed the presence of Y chromosome markers SY620 and AMG, and the absence of SY143, SY254 and SY147. CGH results showed gain of Yq11.2-pter and loss of Yq11.2-q12. These results show the presence of a Yp isodicentric: idic(Y)(q11.2). Case 3 was a 45,X,inv(9)(p11q12)[30]/46,X,idic(Y)(p11.3?),inv(9)(p11q12)[70] karyotype. The FISH signal covered all the abnormal Y chromosome using a Y chromosome paint. The PCR study showed the presence of Y chromosome markers AMG, SY620, SY143, SY254 and SY147. CGH only showed gain of Yq11.2-qter. These results support the presence of an unbalanced (Y;Y) translocation. Our results show that the combined use of molecular and classical cytogenetic methods in clinical diagnosis may allow a better delineation of the chromosome regions implicated in specific clinical disorders.     

Tetrasomy 12p--unusual presentation in CVS

CVS direct preparations usually achieve limited resolution and are better at detecting numerical rather than structural abnormalities. A CVS direct preparation analyzed using G-banding revealed a 47,XY,+G karyotype in 5 of 11 cells and was reported as mosaic for trisomy 21. Subsequent analysis of the CVS culture found only normal male cells. Amniocentesis revealed both normal male cells and cells with an extra F-group chromosome. Fluorescence in situ hybridization (FISH) identified this chromosome to be an isochromosome from the short arm of chromosome 12 [i(12)(p10)]. The amniocyte karyotype was reported as 47,XY,+i(12)(p10)[12]/46,XY[8].ish i(12)(p10)(wcp12+), which is associated with Pallister-Killian syndrome. Reexamination of the CVS direct preparation by FISH with a chromosome 12 centromere probe confirmed the karyotype of this tissue to be 47,XY,+mar[5]/46,XY[6].nuc ish 12cen(D12Z3 x 3)/12cen(D12Z3 x 2). Thus, multiple studies, including amniocentesis and fluorescence in situ hybridization, may be required to fully and accurately evaluate abnormalities detected by CVS. This case also indicates that mosaicism for supernumerary isochromosomes may have a complex origin.