正常人240个人精子染色体核型分析

应用人精子与去透明带金黄地鼠卵受精技术制备人精子染色体标本，对正常男性２４０个精子核型进行了分析。结果显示，数目畸变精子率为３．３４％，结构畸变精子率为６．６７％。所观察到的结构畸变类型为无着丝粒断片，染色体断裂、双着丝粒体、易位和染色单体断裂。在正常男性精子染色体组中，Ｘ一精子与Ｙ一精子的比例为４９．１７％：５０．８３％；近端着丝粒染色体随体联合显随机分布，随体联合频率为０．０９６。对上述实验结果的意义进行了讨论。     

多色荧光原位杂交检测弱精子症的非整倍体

背景与目的:以多色荧光原位杂交技术评价弱精子症患者的精子非整倍体.材料与方法:应用X、Y、18号染色体探针对6例弱精子症患者和3例健康男性进行多色荧光原位杂交试验,检测精子的非整倍体.结果:计数30 305个精子,杂交率99.98%.缺体类型主要为XX18、YY18、XY18、X1818、Y1818.实验组缺体率分别为0.133%±0.095%、0.135%±0.231%、0.438%±0.415%、0.507%±0.973%及0.432%±0.705%,对照组缺体率分别为0.011%±0.020%、0.006%±0.010%、0.051%±0.075%、0.023%±0.040%及0.034%±0.034%.实验组的非整倍体率为2.02%,对照组为0.25%,二者比较,差异具有统计学意义(P＜0.01).结论:多色荧光原位杂交技术对于准确检测人精子非整倍体具有较高的应用价值.     

地鼠血清培养异合卵制备人精子染色体的研究

本文首次应用地鼠血清培养异合卵制备人精子染色体标本，获得显著的效果，同时应用胎牛血清进行了比较，在两咱血清中精子染色体形成率分别为１０２．４５％和５８．２８％，可供分析核型获得率分别为７６．１５％和３１．５１％，两组间存在非常显著的差异（Ｐ〈０．００１）。观察分析了６例供精者的４９８个精子核型，数目异常和结构异常发生率分别为０．８％和２．８％。本研究提示地鼠表培养同种属可促进卵细胞在体外在进一步成     

平阳霉素诱发人精子染色体结构畸变类型

人精子经平阳霉素处理后与去透明带地鼠卵受精，继而制备染色体进行核型分析．结果显示断裂是诱发畸变的主要类型，其后依次为互换、缺失、环和粉碎化；染色体型畸变的比例和均数远高于染色单体型；断裂型畸变的比例和均数远高于重接型．畸变中１７．３％为重接型，表明金黄地鼠卵母细胞中的ＤＮＡ损伤修复系统能够修复化学诱变所致人精子ＤＮＡ损伤．染色体型互换多于染色单体型互换提示：对这类损伤的修复，复制前修复系统可能比复制后修复系统作用更强．研究结果还证实染色体畸变精子和正常精子同样具有与卵受精的能力，提示人精子若反复或是期暴露于环境诱变因子，所产生的ＤＮＡ损伤有可能在精子中积累并在受精时不受选择性淘汰而传递给下一代．     

乙型肝炎病毒对精子染色体的影响

乙型肝炎病毒感染者的精子与去透明带金黄地鼠卵受精制备染色体。并以ＨＢＶ全长ＤＮＡ作探针对精子染色体进行荧光原位杂交。结果发现慢性迁延型肝炎患者,慢性活动型肝炎患者ＨＢＶ携带者的精子染色体畸变率分别为１２．３３％（０／７３）、１６．１８％（１１／６８）和１５．４８％（１３／４８）,均显著高于健康对照者精子畸变率（４．３５％,５／１１５）,进一步ＦＩＳＨ显示,在１例慢性迁延型肝炎患者的精子染色体上有待     

伴der(Y)t(Y;1)(q12;q21)真性红细胞增多症一例并文献复习

目的 探讨伴der(Y)t(Y;1)(q12;q21)真性红细胞增多症(PV)患者的临床特征.方法 分析烟台毓璜顶医院收治的1例伴der(Y)t(Y;1) (q12;q21)PV患者资料,并进行文献复习.结果 该患者诊断为PV,初诊时染色体核型为46,XY,病程13年,白细胞明显增多,染色体核型转变为46,X,der(Y)t(Y;1) (q12;q21).结论 der(Y)t(Y;1)是一种比较罕见的血液系统肿瘤相关的重现性异常.     

人精子诱变检测中新体外代谢活化系统的研究

人精子与去透明带金黄地鼠卵受精,在受粗后异合卵培养阶段,用终浓度分别为２０μｇ／ｍｌ、４０μｇ／ｍｌ、４０μｇ／ｍｌ、６０μｇ／ｍｌ的间接诱变剂环磷酰胺进行处理,继而制备精子染色体进行核型分析。得到环磷酰胺诱发的染色体畸变精子率在上述剂量组依次为３３％、５０％、３３％;断裂均数依次为０．５４、１．９４和０．７２。与空白对照组比较,差异具有显著性。本研究结果表明,用人精子／金黄地鼠异合卵为靶细胞,能     

Ph阴性伴Y染色体缺失的不典型慢性粒细胞白血病一例并文献复习

目的 对一例Ph阴性、bcr-abl融合基因阴性伴Y染色体缺失的不典型慢性粒细胞白血病(CML)患者进行荧光原位杂交(FISH)分析,以检测其是否存在隐蔽的bcr-abl基因重排,排除bcr-abl融合基因假阴性可能,验证其Y染色体缺失的异常核型.方法 采用位点特异性探针GLPbcr-abl(绿/红)额外信号(ES)探针检测bcr-abl基因重排;采用着丝粒探针CSP 18/CSP X/CSP Y(蓝/绿/红)检测Y染色体缺失.结果 GLP bcr-abl ES探针计数200个间期细胞,100%为两红、两绿信号,Ph染色体为阴性;着丝粒CSP 18/CSPX/CSP Y探针计数200个间期细胞,67%细胞为两蓝一绿信号,色信号丢失,存在Y染色体缺失.结合其病情特点,Y染色体缺失为后天获得性.该例Ph阴性、bcr-abl融合基因阴性伴Y染色体缺失的不典型CML患者预后差.结论 Ph阴性、bcr-abl融合基因阴性伴Y染色体缺失的不典型CML患者病情进展凶险,预后差;FISH技术检测染色体异常特异、可靠,有助于对不典型CML患者确诊,关于不典型CML患者的分子生物学本质有待进一步病例积累及研究揭示.     

Ph阴性伴Y染色体缺失的不典型慢性粒细胞白血病一例并文献复习

目的 对一例Ph阴性、bcr-abl融合基因阴性伴Y染色体缺失的不典型慢性粒细胞白血病(CML)患者进行荧光原位杂交(FISH)分析,以检测其是否存在隐蔽的bcr-abl基因重排,排除bcr-abl融合基因假阴性可能,验证其Y染色体缺失的异常核型.方法 采用位点特异性探针GLPbcr-abl(绿/红)额外信号(ES)探针检测bcr-abl基因重排;采用着丝粒探针CSP 18/CSP X/CSP Y(蓝/绿/红)检测Y染色体缺失.结果 GLP bcr-abl ES探针计数200个间期细胞,100%为两红、两绿信号,Ph染色体为阴性;着丝粒CSP 18/CSPX/CSP Y探针计数200个间期细胞,67%细胞为两蓝一绿信号,色信号丢失,存在Y染色体缺失.结合其病情特点,Y染色体缺失为后天获得性.该例Ph阴性、bcr-abl融合基因阴性伴Y染色体缺失的不典型CML患者预后差.结论 Ph阴性、bcr-abl融合基因阴性伴Y染色体缺失的不典型CML患者病情进展凶险,预后差;FISH技术检测染色体异常特异、可靠,有助于对不典型CML患者确诊,关于不典型CML患者的分子生物学本质有待进一步病例积累及研究揭示.     

Ph阴性伴Y染色体缺失的不典型慢性粒细胞白血病一例并文献复习

 目的　对一例Ph阴性、bcr-abl融合基因阴性伴Y染色体缺失的不典型慢性粒细胞白血病（CML）患者进行荧光原位杂交（FISH）分析，以检测其是否存在隐蔽的bcr-abl基因重排，排除bcr-abl融合基因假阴性可能，验证其Y染色体缺失的异常核型。方法　采用位点特异性探针GLP bcr-abl（绿／红）额外信号（ES）探针检测bcr-abl基因重排；采用着丝粒探针CSP 18／CSP X／CSP Y（蓝／绿／红）检测Y染色体缺失。结果　GLP bcr-abl ES探针计数200个间期细胞，100 %为两红、两绿信号，Ph染色体为阴性；着丝粒CSP 18／CSP X／CSP Y探针计数200个间期细胞，67 %细胞为两蓝一绿信号，红色信号丢失，存在Y染色体缺失。结合其病情特点，Y染色体缺失为后天获得性。该例Ph阴性、bcr-abl融合基因阴性伴Y染色体缺失的不典型CML患者预后差。结论　Ph阴性、bcr-abl融合基因阴性伴Y染色体缺失的不典型CML患者病情进展凶险，预后差；FISH技术检测染色体异常特异、可靠，有助于对不典型CML患者确诊，关于不典型CML患者的分子生物学本质有待进一步病例积累及研究揭示。     

骨髓增生异常综合征T淋巴细胞异常与克隆造血

 目的　了解T淋巴细胞异常在骨髓增生异常综合征（MDS）克隆造血中的作用。方法　对76例MDS患者的染色体核型、T淋巴细胞亚群及激活状态进行分析。结果　正常核型36例,异常核型40例,异常发生率52.6 %。40例异常核型中,三体8（+8）24例,占异常核型的60.0 %。与健康对照组比较,MDS患者CD+3 CD-19、CD+3 CD-4 CD+8以及CD+3 HLA-DR+细胞百分率显著升高,CD-3（CD16 CD56）+细胞的百分率明显降低。将MDS患者进行核型分组,异常核型组CD+3（CD16 CD56）+细胞的百分率显著高于正常对照组。将+8核型从MDS异常核型中独立出来进行分析,CD+3 CD+4 CD-8细胞的百分率明显低于正常核型以及其他异常组,CD4／CD8的比值明显低于健康对照组。结论　MDS存在T淋巴细胞异常,异常核型MDS可能恶性克隆增殖更为优势,预后更差。+8 核型MDS存在更为严重的免疫监视功能下降,导致恶性克隆过度增殖与残存造血过度受抑。     

骨髓增生异常综合征124例细胞遗传学变化与预后的关系

 【摘要】　目的　探讨骨髓增生异常综合征（MDS）细胞遗传学的变化与预后的关系。方法　回顾性分析2005年9月至2009年10月确诊的124例骨髓增生异常综合征患者,男79例,女45例,年龄14～79岁,中位年龄 57岁。难治性贫血（RA）26例,难治性贫血伴铁粒幼细胞增多型（RAS）13例,难治性贫血伴有多系病态造血（RCMD）21例,难治性贫血伴有幼稚细胞增多Ⅰ型（RAEB-Ⅰ）29例,难治性贫血伴有幼稚细胞增多Ⅱ型（RAEB-Ⅱ）35例 。124例患者应用细胞短期培养法及热处理吉姆萨反带技术,分析20个分裂期细胞,随访至2011年12月,根据患者染色体核型情况,分析MDS各个亚型正常核型组和异常核型组,3年转化成白血病发生率（转白率）及生存率。结果　39例RA 、RAS患者中正常核型32例,异常核型7例,核型异常率17.9 %,而在85例RCMD、RAEB-Ⅰ、RAEB-Ⅱ患者中,检测出异常核型38例,核型异常率44.4 %,较RA、RAS明显增高32例,RA、RAS正常核型有2例3年后转化为白血病,7例异常核型中有1例3年后转化为急性白血病。而在38例RCMD、RAEB-Ⅰ、RAEB-Ⅱ异常核型中,随访3年,有16例转化为白血病,转白率42.1 %（16／38）,47例正常核型中有13例转化为白血病,转白率为27.6 %（13／47）,两组比较差异有统计学意义（P＜0.05）,RCMD、RAEB-Ⅰ、RAEB-Ⅱ组转白率较RA、RAS明显增高,尤其在核型异常组转白率高达42.1 %。结论　MDS伴有异常核型者,转白率高,治疗效果差,宜早期行造血干细胞移植治疗。     

Intensive chemotherapy is not recommended for patients aged >60 years who have myelodysplastic syndromes or acute myeloid leukemia with high-risk karyotypes

BACKGROUND: It is unclear whether intensive chemotherapy is beneficial to patients with high-risk myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML) if they are aged >/=60 years. METHODS: The authors studied 160 patients with a median age of 67 years who received intensive chemotherapy for MDS or AML with cytosine arabinoside and an anthracycline. RESULTS: At diagnosis, cytogenetic analysis was available in 146 patients. Karyotype was normal in 78 patients and abnormal in 68 patients. Of the abnormal karyotypes, 32 belonged to the high-risk category, ie, they involved either >/=3 chromosomes or chromosome 7. Complete remission (CR) was achieved by 94 patients (56%). CR rates were 70% among the patients who had a normal karyotype, 69% among the patients who had an abnormal (noncomplex) karyotype, but only 46% among the patients ho had a high-risk karyotype. The median survival was 9.5 months in the entire group, 18 months in patients with normal karyotype, 6 months in patients with abnormal, and 4 months in patients with a high-risk karyotype. A poor prognosis was attributable to low rates of CR and a high risk of early recurrence. CONCLUSIONS: According to the current data, elderly patients with AML or advanced MDS do not benefit from intensive chemotherapy if they show karyotype anomalies, especially those in the high-risk category.     

Novel mutations of the nucleophosmin (NPM-1) gene in Egyptian patients with acute myeloid leukemia: A pilot study

Mutations of the nucleophosmin (NPM-1) gene have been reported in 50-60% of acute myeloid leukemia (AML) patients with normal karyotype. This work was designed to study the prevalence and nature of NPM1 gene mutations in a group of Egyptian patients with AML to get an idea about the profile of NPM1 gene mutations in our society. In 45 previously untreated patients with de novo AML, peripheral blood and/or bone marrow samples from all patients were subjected to microscopic morphologic examination, cytochemical analysis, immunophenotyping and karyotyping. Patients with normal cytogenetic results were selected for molecular analysis of NPM1 exon 12 by PCR amplification followed by DNA sequencing of the amplified product. Twenty-one patients (46.7%) had abnormal karyotype: six cases with t(15;17), five cases with t(8;21), five cases had trisomy 8, two cases carrying inv(3) and three cases had monosomy 7. The remaining 24 patients (53.3%) had normal karyotype. These patients were then subjected to molecular analysis. Out of these 24 patients with normal karyotype, mutant NPM-1 was detected in 11 patients (45.8%) by DNA sequencing; 2 cases showed type A mutation, 2 cases were harboring [ins 1015-1019 (CACG)], with point mutation [1006C>G], while the remaining 7 cases showed heterozygous deletion of nt A [del 1178 (A)]. Conclusion: Two novel NPM1 gene mutations were detected among our study population of AML patients identified as: the insertion CACG associated with point mutation, deletion of one base, or associated with point mutation. NPM1 gene mutations may become a new tool for monitoring minimal residual disease in AML with normal karyotype. Whether these previously unreported NPM-1 mutations will confer the same better outcome as previously reported mutations is currently unknown and warrants a larger study. Conclusion: Two novel NPM1 gene mutations were detected among our study population of AML patients identified as: the insertion CACG associated with point mutation, deletion of one base, or associated with point mutation. NPM1 gene mutations may become a new tool for monitoring minimal residual disease in AML with normal karyotype. Whether these previously unreported NPM-1 mutations will confer the same better outcome as previously reported mutations is currently unknown and warrants a larger study.     

FISH analysis in addition to G-band karyotyping: Utility in evaluation of myelodysplastic syndromes?

Cytogenetic analysis provides important diagnostic and prognostic information for patients with myelodysplastic syndromes (MDS). Prior studies, mostly comprised of small sample sizes, have reported conflicting results while evaluating the usefulness of FISH in addition to G-band karyotyping in MDS. In the current study, the utility of performing a tailored FISH panel, in addition to G-band karyotyping was evaluated in a series of 110 MDS patients diagnosed at our institute. Using our FISH panel, clonal cytogenetic abnormalities were detected in 3/8 (38%) of MDS cases with karyotype failure and in 5/54 (9%) cases with normal G-band karyotypes, all the latter had intermediate or high grade MDS. Of the cases with abnormal G-band karyotypes, 6/48 (13%) showed discrepancies between FISH and G-band results, however, FISH analysis only lead to reassignment of karyotypic abnormalities to different chromosomes, MDS cytogenetic risk stratification was not altered. Our findings suggest that FISH testing is informative only in MDS cases with karyotype failure and intermediate-high grade MDS cases with normal G-band karyotype and has limited utility in cases that have normal G-band karyotypes and morphologic features of low grade MDS or in cases with abnormal G-band karyotypes.     

Ovarian and testicular damage by cytotoxic drugs.

The use of cytotoxic drugs has improved the survival figures in malignancies. These drugs exert harmful effects on the tumour cell and normal cells as well. The detrimental effects on gonads were studied in 36 patients who had cytotoxic chemotherapy for various malignancies. In Group I (20 adults in clinical remission after chemotherapy), sterility was noted in 93.7% of men and hundred percent of women experienced irregularly mensus during treatment, 50% of them subsequently developing persistent amenorrhoea. In Group II, (16 adults undergoing chemotherapy) 100% of men had oligospermia, with 75% of them developing azoospermia after 2-3 cycles of treatment. Women also showed similar effect, but were less vulnerable than men.     

异常核型骨髓增生异常综合征64例预后分析

 【摘要】　目的　探讨骨髓增生异常综合征（MDS）患者染色体异常与预后的关系,对治疗效果进行分析。方法　回顾性分析122例MDS患者染色体核型,用吉姆萨显带法进行检测。难治性贫血（RA）、环形铁幼粒细胞难治性贫血（RAS）的治疗以诱导分化剂及刺激造血药物为主。原始细胞过多难治性贫血（RAEB）、转化型原始细胞过多难治性贫血（RAEB-t）、慢性粒-单核细胞白血病（CMML）的治疗以小剂量化疗和小剂量联合化疗方案为主。分析异常核型MDS患者疗效,以同期住院的正常核型MDS患者为对照。结果　检出异常核型MDS患者64例,治疗后完全缓解（CR）17例,CR率26.6 %。同期正常核型MDS患者58例,CR 30例,CR率51.7 %。正常和异常核型患者CR率差异有统计学意义（χ2＝8.13,P＝0.04）。复杂核型、-7、+8核型异常者易进展为急性白血病。结论　染色体核型分析在MDS的诊断与预后判断中有重要意义,不同的染色体核型改变进展为白血病的风险不同。     

Cytogenetic progression and prognosis in oral-carcinoma - a DNA flow cytometric study on 317 cases

A total number of 317 consecutive patients with squamous cell carcinoma of the oral cavity were analysed by DNA flow cytometry. The proportion of tumors consisting exclusively of diploid cells decreased from 75% in T1G1 carcinomas to less than 10% in advanced tumors in favor of cases characterized by the presence of cell lines with aberrant DNA content. This observation indicates a genetic evolution of tumor clones with severe karyotype aberrations from diploid progenitors which obviously represents an ultimate event in the pathogenesis of oral carcinomas. Aneuploidy formation clearly contributes to the acquisition of progressive malignant behaviour as is underlined by a dramatic difference of the survival outcome of patients with diploid tumors (5-year overall survival: 88%) as compared to those who had aneuploid tumors (36%; P<0.001), a relationship which held true even if identical tumor stages were considered. There were no indications of successive genetic changes of the cellular DNA contents during tumor progression which would explain the wide range of variation of the individual DNA values. This observation supports the hypothesis of an aneuploidy formation by a single cytogenetic event, such as an abnormal mitosis. Although there were indications of an even worse survival outcome in the group of patients with peri-diploid tumor cell lines (P<0.1), a significant influence of different DNA contents on the prognosis could not be detected.     

急性淋巴细胞白血病80例染色体核型分析及其与疗效的相关性

目的 探讨急性淋巴细胞白血病（ALL）患者的染色体核型特点及其与疗效的关系.方法 采用短期培养法、R显带技术对80例ALL患者骨髓染色体核型进行分析.结果 80例ALL患者中,正常核型53例（66.2％）,异常核型27例（33.8％）,其中,染色体核型结构异常者10例（12.5％）,数目异常者2例（2.5％）,复杂异常15例（18.8％）.按数目畸变分类,＞ 50的超二倍体2例（2.5％）、47～50的超二倍体5例（6.25％）、假二倍体18例（22.5％）、正常二倍体53例（66.25％）、亚二倍体2例（2.5％）、未见近三倍体或近四倍体.正常核型患者疗效优于异常核型患者（x2=19.371,P＜ 0.01）,复杂核型患者疗效差于其他核型患者（x2=9.145,P=0.004）,伴有t（9;22）（q34;q11）患者疗效差于其他核型患者（x2=5.785,P=0.021）.结论 ALL患者的染色体核型异常具有随机性,常见的异常核型为复杂核型和伴有t（9;22）（q34;q11）核型,其疗效均较差.     

儿童急性淋巴细胞白血病遗传学特征

目的：分析儿童急性淋巴细胞白血病（ALL）遗传学变化的特点.方法：以94例初发ALL患儿为研究对象,采用染色体核型分析及多重PCR技术检测患者骨髓白血病细胞的细胞遗传学和分子生物学变化.结果：63例患儿做了核型分析,有分裂象者55例,其中异常者12例（19.0%）;无分裂象者8例（12.7%）.对93例患儿进行了多重PCR检测,其中包括53例染色体有分裂象患儿和8例无分裂象的患儿,41例正常染色体核型患儿中异常者为17例（占41.5%）,12例染色体核型异常者中融合基因异常者4例（33.3%）,无分裂象患者异常2例,无分裂象检出率与有分裂象者核型分析异常检出率差异无统计学意义;93例患儿中融合基因检出率为34.4%（32/93）,其中TEL/AML1融合基因阳性20例（21.51%）,MLL重排3例（3.23%）,BCR/ABL（p190）融合基因阳性3例（3.23%）,E2A/PBX阳性5例（5.38%）,HOX11阳性1例.94例患儿通过两种检测方法共检出细胞遗传学异常46例（48.94%）.结论：儿童ALL可出现细胞遗传学改变.对于有足够分裂象的患者,常规染色体核型分析仍是可靠的方法.对分裂象质量低或无分裂象的儿童ALL患者,多重PCR检测异常克隆能提高细胞遗传学异常检出率.将染色体核型分析及多重PCR结合起来可提高细胞遗传学异常检出率.