男性胃癌病人Y染色体缺乏的原位杂交检测及其意义

建立了用染色体特异的探针检测细胞染色体畸变的原位杂交法。用生物素标记的ＰＹ３．４探针对１２例男性胃癌标本进行原位杂交，检测Ｙ染色体的丢失，我们发现：胃癌组织细胞的Ｙ染色体丢失率很高，Ｙ染色体丢失很可能是胃癌细胞的一个染色体标志，胃癌组织细胞的Ｙ染色体丢失与肿瘤的分化程度和肿瘤的转移有一定关系，这对胃癌的诊断和估计预后有一定帮助。     

人类Y染色体特异DNA探针(PY3.4)的染色体原位杂交

DNA重组技术与染色体分析相结合,是深入研究染色体微小变异的有力手段。本实验采用人类γ染色体特异DNA探针(PY 3.4)进行细胞和染色体原位杂交。 把含有人类Y染色体特异DNA片段(Y 3.4)的重组质粒扩增提取后,用EcoRI酶切,DE-81滤纸回收,按照缺口翻译标记,标记前体为~3HdTTP和~3HdATP,经Sephadex G-50柱层析分离,得到     

男性胃癌病人Y染色体缺失的原位杂交检测及其意义

建立了用染色体特异的探针检测细胞染色体畸变的原位杂交法。用生物素标记的ＰＹ３．４探针对１２例男性胃癌标本进行原位杂交，检测Ｙ染色体的丢失，我们发现：胃癌组织细胞的Ｙ染色体丢失率很高，Ｙ染色体丢失很可能是胃癌细胞的一个染色体标志，胃癌组织细胞的Ｙ染色体丢失与肿瘤的分化程度和肿瘤的转移有一定关系，这对胃癌的诊断和估计预后有一定帮助。     

应用直接酪胺信号放大方法显示Y染色体荧光原位杂交信号

目的用直接酪胺信号放大(tyramine signal amplification,TSA)方法显示Y染色体荧光原位杂交信号,以提高检测脑组织切片Y染色体荧光原位杂交信号的灵敏度。方法以小鼠Y染色体重复序列pY353/B cDNA为模板标记RNA探针,以雌性小鼠为对照,用原位杂交方法杂交Y染色体特异性基因区域,分别用直接TSA方法和"三步抗体法"显示荧光原位杂交信号。通过计数Y染色体阳性细胞核占所有细胞核的百分率来计算TSA法和"三步抗体法"检测脑组织切片Y染色体荧光原位杂交信号的灵敏度,比较分析两种方法灵敏度的差异。结果"三步抗体法"在脑切片检测Y染色体阳性信号的灵敏度和特异度分别为85.67%和100%;直接TSA法在脑切片检测Y染色体阳性信号的灵敏度和特异度分别为92.82%和100%。结论TSA方法检测脑组织切片Y染色体荧光原位杂交信号的灵敏度比"三步抗体"法有显著的提高,可以用于脑内追踪和分析骨髓干细胞的迁移规律。     

双色荧光原位杂交检测男性不育患者精子X和Y染色体

目的建立男性不育患者精子核双色荧光原位杂交技术（FISH）的检测方法。方法将20例男性不育患者精子标本经低渗液处理、固定后制片,用二硫苏糖醇（DTT）使精子去凝集,变性后与双色荧光标记（CEPX、Y）探针杂交。结果20例患者的6672个精子中,X染色体精子率为47．78％,Y染色体精子率为47．24％,性染色体数目异常的精子率为4．53％,有效杂交率为99．55％。结论成功地建立了检测人精子X和Y染色体的检测方法。     

大Y染色体的遗传学探讨

目的：分析大Y染色体与各种临床表现的相关性。方法：采用外周血淋巴细胞G显带技术，对患者进行核型分析。结果：41例大Y染色体患者中，临床表现有：其妻有异常生育史，有睾丸发育不全者，无精和精子异常者，有智力低下儿和孤独症儿等。结论：大Y染色体与临床疾病有一定的相关性。     

利用PCR，FISH对异常核型中额外染色体的证实

目的：检测在常规G显带下，45，X［115］/46，X+mar［45］/46，XY［29］异常核型中额外小染色体的来源。方法：利用PCR对SRY基因进行检测；对人的Y染色体进行标记，利用荧光原位杂交技术，在荧光显微镜下观察。结果：用Y染色体杂交，发现在额外小染色体上有荧光信号。结论：额外小染色体来源于Y染色体。     

荧光原位杂交法检测特纳氏综合征患者的微小标记染色体

目的：确定特纳氏综合征（Turner‘s syndrom)患者微小标记染色体（Small marker chromosome,smr)的来源。方法：用X和Y染色体着丝粒特异的DNA探针进行荧光原位杂交（Fluorescence in situ hybridization,FISH)，检测3例核型为45，X/46，X smr的特纳氏综合征患者的smr。结果：2例患者smr染色体来源于Y染色体，1例来源于X染色体。结论：FISH技术可快速准确鉴别微小标记染色体，对临床诊断和治疗方案的选择有重要指导作用。     

生物素标记DNA探针测定人性别

白细胞或胎儿绒毛组织染色体DNA经限制酶Hae (?)消化后,用光生物素(photo-biotin)标记人Y染色体特异的3.4 kb DNA片段作探针,进行Southern印迹杂交,并以生物素结合蛋白-碱性磷酸酶体系显示生物素探针杂交信号。结果表明,0.1μg男性DNA可显示清晰的信号,而5μg女性DNA则无信号出现;平行试验证明生物素探针测定人性别的结果与32P标记探针完全一致。提示生物素探针可用于X连锁遗传病、性异常,运动员体检,法医学及异性别骨髓移植方面的性别分析。     

小Y染色体对男性生育力的影响及临床效应分析

目的 探讨小Y染色体对男性生育能力的影响及临床效应分析。 方法 收集因不孕不育或配偶有不良孕产史就诊的男性患者1 804例(实验组),分为2组,其中A组1 269例,拟行体外授精/卵胞质内单精子注射-胚胎移植(IVF/ICSI-ET)技术; B组535例,配偶既往均有不良孕产史或羊水穿刺提示胎儿染色体异常等。对照组为行羊水穿刺诊断结果为正常的男性病例656例。采用染色体G显带技术对两组患者进行核型分析,筛选小Y染色体。对检出的小Y患者应用PCR荧光探针法检测Y染色体无精子因子(AZF)的相关序列标签位点。 结果 实验组共检出小Y染色体44例,其中A组31例,B组13例(已剔除1例); 对照组检出6 例。A组及B组的小Y染色体检出率与对照组比较,差别均有统计学意义(P<0.05),而A组与B组间差别无统计学意义(P>0.05)。Y染色体微缺失检测发现,A组AZF微缺失3例,其中2例为AZFc缺失,1例为AZFb+c复合缺失,B组未发现AZF微缺失,AZF微缺失发生率为6.81%(3/44)。 结论 小Y染色体可能与男性不育及不良妊娠结局的发生有关,存在潜在的临床效应。     

用Y特异DNA探针对染色体核型46,X,t(X;Y)(p22.3;q12.1)患者的分析

本文报道一女性的第一、二性征均正常,但GTG显带的核型中可见到下条异常X染色体短臂增长,断裂点为Xp22.3,CBG显带核型中异常X染色体均连接1个额外浅染片段。用凝胶原位杂交法以Y特异DNA探针进行DNA分析,分子杂交图谱与正常男性相一致,证实患者基因组DNA含有DYZ_1重复顺序。故该女性染色体核型为46,X,t(X; Y) (qter→p22.3::q12.1→qter).此外,对患者的临床特征、女性性腺激素、染色体异常产生原因及有关问题作了讨论。     

Molecular genetic evidence of Y chromosome loss in male patients with hematological disorders

Background There has been continuous debate as to whether Y chromosome loss is an age related phenomenon or a cytogenetic marker indicating a malignant change. This study aimed to investigate the frequency of Y chromosome loss in the specific patients in order to determine whether it is an age related phenomena or a cytogenetic marker indicating a malignant change. Methods Five hundred and ninety-two male patients with a median age of 59 years old (22–95 years) were included in this study. These patients were divided into two groups: the study group, including 237 patients who had hematological disorders included myeloproliferative disorder (MPD), myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), multiple myeloma (MM), and lymphoma and the control group including 355 patients with no evidence of hematological disease. Both conventional cytogenetics and fluorescence in situ hybridization using DNA probes specific for the centromere of chromosomes X or Y were performed according to our standard laboratory protocols.Results Twenty-four out of 237 patients with hematological disorders (10.1%) had Y chromosome loss. Of these 24 patients, 2 patients had AML (5.0% of all AML patients), 2 patients had CML (5.7% of all CML patients), 2 patients had MPD (8.0% of all MPD patients), 3 patients had MM (10.0% of all MM patients), 5 patients had lymphoma (10.6% of all lymphoma patients) and 10 patients had MDS (16.7% of all MDS patients). Twenty-one out of these 24 patients had a loss of Y chromosome as the sole anomaly and the remaining three had a loss of Y chromosome accompanied with other structural changes detected by conventional cytogenetic analysis. Fluorescence in situ hybridization (FISH) analysis confirmed the routine cytogenetic results. All 24 patients had a loss of Y chromosome with a range of 17.5%–98.5% of cells. Two of the patients, one with AML and another with CML, had karyotype and FISH testing done both at the initial diagnosis and during remission. The results showed a loss of Y chromosome at initial diagnosis but a normal 46,XY karyotype during remission. Only 9 out of 355 patients (2.5%) without evidence of hematological disease had Y chromosome loss, among them 7 patients had cardiovascular diseases and 2 patients had kidney diseases. Comparison of the incidence of Y chromosome loss in patients with hematological disorders or without evidence of hematological disease using statistical analysis showed a statistically significance difference (P&lt;0.05).Conclusions The present study demonstrated that the frequency of Y chromosome loss is significantly higher in patients with hematological disorders than in patients without hematological disorders, which indicates that the loss of Y chromosome is associated with a neoplastic change.     

用Y染色体特异的DNA探针对冷冻异体胎儿真皮移植后的追踪观察

为了直观而特异地观察了追踪冷冻异体胎儿真皮移植后的转归。采用生物素标记的人Ｙ染色体特异的ＤＮＱＡ探针，对５例临床冷冻异体异性胎儿真皮－自体表皮复合移植患者的６个标本作原位杂交。异体胎儿真皮移植１２，１９，２０，２１和４１ｄ后仍存活。证明冷冻异体胎儿真皮复合移植后至少可存活４１ｄ。     

Y染色体微缺失与男性不育相关

目的：探讨Y染色体微缺失原因不明性无精症、少精症所致男性不育的相关性。方法；特发性少精症18例、特发性严重少精症12例，特发性无精症5例及正常已生育健康男性10例作为研究对象和正常对照，应用多重PCR技术，对精液基因组DNA进入Y染色体上连续的18个序列标记位点检测。结果：少精症18例中发现Y染色体微缺失2例，严重少精症12例中发现Y染色体微缺失3例，无精症5例和正常已生育男性10例均末发现Y染色体微缺失，缺失形式3种。结论：Y染色体微缺失与精子发生障碍相关。     

Identification of cultured male epidermal allografts by detecting Y chromosome DNA fragments in female burn victims

ltiswellknownthattheskinofextensivelyburnedcasesthemselvesisinsufficienttoprovidecoverageforwounds.Thoughallogeneicskincancoverthewoundstemporari1y,itwillberejectedwithin2-3weeks.Thiscanthreatenthepatient'slifeifthereisnoenoughautogenousskintoreplacetherejectedoneontime.GreenetalL1]reportedthattheburnwoundsofpatientswithextensiveburnscouldbetreatedbytransplantingepidermalautograftsafterautologousepidermalcellsmulti-plyforthousandsoftimesinvltrowithinashortperiodoftime-Butittakesatleast2-3weeks…