Y chromosome microdeletions in azoospermic patients with Klinefelter＇s syndrome

Aim： To study the occurrence of Y chromosome microdeletions in azoospermic patients with Klinefelter＇s syndrome （KFS）. Methods： Blood and semen samples were collected from azoospermic patients with KFS （n = 14） and a control group of men of proven fertility （n = 13）. Semen analysis was done according to World Health Organization （WHO） guidelines. Blood samples were processed for karyotyping, fluorescent in situ hybridization （FISH） and measurement of plasma follicle stimulating hormone （FSH） by radioimmunoassay. To determine Y chromosome microdeletions, polymerase chain reaction （PCR） of 16 sequence tagged sites （STS） and three genes （DFFRY, XKRY and RBM1 Y） was performed on isolated genomic DNA. Testicular fine needle aspiration cytology （FNAC） was done in selected cases. Results： Y chromosome microdeletions spanning the azoospermia factor （AZF）a and AZFb loci were found in four of the 14 azoospermic patients with KFS. Karyotype and FISH analysis revealed that, of the four cases showing Y chromosome microdeletion, three cases had a 47,XXY/46,XY chromosomal pattern and one case had a 46,XY/47,XXY/48,XXXY/48,XXYY chromosomal pattern. The testicular FNAC of one sample with Y chromosome microdeletion revealed Sertoli cell-only type of morphology. However, no Y chromosome microdeletions were observed in any of the 13 fertile men. All patients with KFS had elevated plasma FSH levels. Conclusion： Patients with KFS may harbor Y chromosome microdeletions and screening for these should be a part of their diagnostic work-up, particularly in those considering assisted reproductive techniques. （Asian JAndrol 2006 Jan; 8： 81-88）     

Y chromosome microdeletions in azoospermic patients with Klinefelter's syndrome

Aim:To study the occurrence of Y chromosome microdeletions in azoospermic patients with Klinefelter's syndrome(KFS).Methods:Blood and semen samples were collected from azoospermic patients with KFS(n=14)and acontrol group of men of proven fertility(n=13).Semen analysis was done according to World Health Organization(WHO)guidelines.Blood samples were processed for karyotyping,fluorescent in situ hybridization(FISH)andmeasurement of plasma follicle stimulating hormone(FSH)by radioimmunoassay.To determine Y chromosomemicrodeletions,polymerase chain reaction(PCR)of 16 sequence tagged sites(STS)and three genes(DFFRY,XKRYand RBM1 Y)was performed on isolated genomic DNA.Testicular fine needle aspiration cytology(FNAC)was donein selected cases.Results:Y chromosome microdeletions spanning the azoospermia factor(AZF)a and AZFb lociwere found in four of the 14 azoospermic patients with KFS.Karyotype and FISH analysis revealed that,of the fourcases showing Y chromosome microdeletion,three cases had a 47,XXY/46,XY chromosomal pattern and one casehad a 46,XY/47,XXY/48,XXXY/48,XXYY chromosomal pattern.The testicular FNAC of one sample with Ychromosome microdeletion revealed Sertoli cell-only type of morphology.However,no Y chromosome microdeletionswere observed in any of the 13 fertile men.All patients with KFS had elevated plasma FSH levels.Conclusion:Patients with KFS may harbor Y chromosome microdeletions and screening for these should be a part of their diagnosticwork-up,particularly in those considering assisted reproductive techniques.(Asian J Androl 2006 Jan;8:81-88)     

Screening for microdeletions on the long arm of chromosome Y in 53 infertile men

About 30% of couple infertilities are of male origin. They appear in some cases de novo and are considered idiopathic. The aim of our work was to evaluate, in these cases, the prevalence of microdeletions of the long arm of chromosome Y, within the AZF a, b and c regions using molecular biology techniques. Men with azoospermia or oligozoospermia resulting from hereditary, endocrine or obstructive causes, or with a constitutional cytogenetic abnormality were excluded. Fifty-three infertile men with azoospermia or oligozoospermia, as determined by a spermiogram, were studied. Of these, 34 were idiopathic and 7 exhibited a past history of genital infection or biological abnormalities, suggesting partial obstruction of the genito-urinary tract. A further 8 men had a varicocele and 11 cases with a history of cryptorchidism were also studied. Peripheral blood DNA was extracted from each patient, then amplified by multiplex PCR with STS genomic markers from the three Y chromosome AZF zones. PCR products were then analysed on agarose gels. In view of the difficulty of confirming the absence of a signal in molecular biology, each case suspected of having a deletion was checked by multiplex PCR through coamplification with the SRY marker. Five men with microdeletions of the long arm of the Y chromosome were diagnosed among the 53 patients. All of them included the AZFc zone and the intragenic DAZ gene markers. Furthermore, a larger Y chromosome deletion encompassing the 3 AZF zones was diagnosed, and confirmed by cytogenetic analysis. All Y chromosome microdeletions were observed in the 34 truly idiopathic azoospermia/oligozoospermia cases, corresponding to a proportion of 14.7% (or 9.4% considering the whole population of 53 infertile men). The relatively high proportion of microdeletions found in our series suggests the need for strict patient selection to avoid unnecessary screening for long arm Y chromosome microdeletions.     

Anesthesia and Prader-Willi syndrome: preliminary experience with regional anesthesia

The constellation of neonatal hypotonia, developmental delay, hypogonadism and obesity caused by hyperphagia was first reported in 1956 and subsequently termed Prader-Willi syndrome (PWS). Genetic analysis has demonstrated abnormalities of chromosome 15. Anesthesia concerns of PWS include morbid obesity, the potential for difficulties with airway management, risk for perioperative respiratory failure, abnormalities in the central control of ventilation and temperature, rare reports of primary myocardial involvement, aggressive and at times violent behavior and glucose intolerance. For the first time, we report the use of regional anesthesia in four patients with PWS. A lumbar plexus catheter was used to provide postoperative analgesia in one patient while regional anesthesia (fasica iliaca block, spinal anesthesia, and lateral vertical infraclavicular block) was used to provide primary intraoperative anesthesia in three other patients while avoiding the need for general anesthesia. Previous reports of the anesthesia care of patients with PWS are reviewed and the potential perioperative implications of the sequelae of PWS are discussed.     

Y microdeletions in the Istria county, Croatia

按照演化经济学的观点,从经济学的角度可以找到企业核心竞争力的真正内涵。首先对企业的核心竞争力理论分别从经济学与管理学方面做简要的阐述,然后利用演化经济学的理论,对企业的核心竞争力——“惯例”作了分析、归纳,并给出了企业惯例的假设与特征。     

Screening for Y chromosome microdeletions in childhood: lack of evidence for a direct association with testicular maldescent

The aim of the study was to investigate the hypothesis that Y chromosome microdeletions are directly implicated in testicular maldescent. Genomic DNA was extracted from the peripheral blood of 292 subjects. This population consisted of (i) 180 children with all phenotypes of isolated (non‐syndromic) testicular maldescent from 174 index families, (ii) affected adult relatives available (n = 12) and (iii) 100 unrelated children with normal external genitalia (controls). The sequence‐tagged site primer set and the conditions of conventional polymerase chain reaction amplification were based on the current laboratory guidelines for molecular diagnosis of Y chromosome microdeletions recommended by the European Academy of Andrology and the European Molecular Genetics Quality Network. Two multiplex reactions were designed to screen the regions of azoospermic factors a, b and c. Each multiplex reaction included adequate internal and external amplification controls. Amplification products were submitted to electrophoresis on 2% agarose gel impregnated with ethidium bromide dye solution for 80 volt‐h and visualised under ultraviolet light. No microdeletions were detected in any subject. These results indicate that Y chromosome microdeletions are not directly implicated in the pathogenesis of testicular maldescent. Other factors should be investigated to potentially explain the genetic predisposition that seems to exist in at least a subgroup of these patients.     

Routine screening for classical azoospermia factor deletions of the Y chromosome in azoospermic patients with Klinefelter syndrome

Aim： To evaluate the occurrence of classical azoospermia factor （AZF） deletions of the Y chromosome as a routine examination in azoospermic subjects with Klinefelter syndrome （KS）. Methods： Blood samples were collected from 95 azoospermic subjects with KS （91 subjects had a 47,XXY karyotype and four subjects had a mosaic 47,XXY/46, XY karyotype） and a control group of 93 fertile men. The values of testosterone, follicle stimulating hormone （FSH） and luteinizing hormone （LH） were measured. To determine the presence of Y chromosome microdeletions, polymerase chain reaction （PCR） of five sequence-tagged site primers （sY84, sY 129, sY 134, sY254, sY255） spanning the AZF region, was performed on isolated genomic DNA. Results： Y chromosome microdeletions were not found in any of the 95 azoosperrnic subjects with KS. In addition, using similar conditions of PCR, no microdeletions were observed in the 93 fertile men evaluated. The level of FSH in KS subjects was higher than that in fertile men （38.2 ± 10.3 mIU/mL vs. 5.4 ±2.9 mIU/mL, P 〈 0.001） and the testosterone level was lower than that in the control group （1.7 ±0.3 ng/mL vs. 4.3 ± 1.3 ng/mL, P 〈 0.001）. Conclusion： Our data and review of the published literature suggest that classical AZF deletions might not play a role in predisposing genetic background for the phenotype of azoospermic KS subjects with a 47,XXY karyotype. In addition, routine screening for the classical AZF deletions might not be required for these subjects. Further studies including partial AZFc deletions （e.g. gr/gr or b2/b3） are necessary to establish other mechanism underlying severe spermatogenesis impairment in KS.     

无精症患者Y染色体微缺失的多重PCR筛查

目的 建立一套Y染色体微缺失的多重PCR筛查方法。方法 建立5套稳定和可靠的多重PCR筛查方法，对进行ICSI治疗的26例无精子症患者和进行睾丸活检的30例无精子症患者进行Y染色体微缺失的检测。结果 在56例无精子症患者中发现5例AZFc／DAZ缺失，2例同时具有AZFc／DAZ和AZFb／RBMl的缺失，1例仅具有sY153一个片段的缺失。结论 本研究Y染色体微缺失的多重PCR筛查方法是易行和可靠的，对无精子症患者有必要进行Y染色体微缺失的常规筛查。     

精索静脉曲张严重少精子症患者Y染色体微缺失分析

目的:探讨精索静脉曲张和Y染色体微缺失对生精障碍的影响。方法:对随机挑选的100例左侧精索静脉曲张严重少精子症患者(精子浓度<5×106/ml,组1),100例左侧精索静脉曲张轻度少精子症患者[精子浓度(10～20)×106/ml,组2],100例特发性严重少精子症患者(组3),100例特发性轻度少精子症患者(组4)和30例正常生育男性对照组(组5)采用聚合酶链反应(PCR)技术进行Y染色体微缺失检测,选取无精子因子(AZF)a、b和c区9个序列标签位点(STS)。结果:组1中有19例患者存在AZF微缺失(19%);组3中有11例患者存在AZF微缺失(11%),其余各组均未发现AZF微缺失;组1比组3有较高的缺失率。结论:在治疗精索静脉曲张严重生精障碍患者前应先进行Y染色体微缺失检测,避免不必要的治疗。     

Molecular study on Y chromosome microdeletions in Egyptian males with idiopathic infertility

Aim:To determine the frequency of genetic deletions within the azoospermia factors in Egyptian infertile males.Methods:The Yq microdeletions in 33 infertile males with undetectable chromosomal anomalies were examined by mutiplex polymerase chain reaction (PCR).Deletions were confirmed using single PCR amplifications.Results:Four out of the total 33 (12%) men had Yqll microdeletions,thus supporting the average reported figures in other populations.Three of those 4 cases had single short tandem sequence deletions with discrete histological findings of their testes.Single sY272 deletion within AZFc was associated with Sertoli cell only syndrome,whereas a patient with isolated sY84 deletion within AZFa had immature testicular structure.The remaining case had a large deletion in AZFa-c and short stature.Conclusion:The present study supports the hypothesis that the Yq11 encompasses genetic determinants of stature besides genes controlling spermatogenesis.(Asian J Androl 2004 Mar;6:53-57)     

精索静脉曲张男性不育患者Y染色体微缺失检测

目的:探讨精索静脉曲张(varicocele,VC)不育患者Y染色体微缺失特点及其与临床表型的关系,为评价VC不育患者是否行手术治疗或ICSI提供依据。方法:VC不育患者174例,分为3组,A组:无精子症47例;B组:严重少精子症57例;C组:轻度少精子症70例;设立正常生育的健康志愿者男性28例作为对照组(D组)。抽取外周血提取DNA,选取Y染色体上AZFa、AZFb、AZFc区共6个序列标签位点,应用多重PCR进行扩增;已生育女性26例作为阴性对照,分别运用琼脂糖凝胶电泳分离,对照阅读扩增产物,判定有无缺失存在以及缺失类型。结果:174例男性不育患者中有22例检测到Y染色体微缺失,缺失率12.64%;A组11例存在微缺失,B组11例存在微缺失,C组未检测到微缺失。A组与C组、B组与C组比较,差异均有显著性。A组缺失病例中有6例为AZFc区缺失,1例为AZFa缺失,2例为AZFb区缺失,2例为AZFb、AZFc区共同缺失;B组缺失病例中有8例为AZFc缺失,2例为AZFb缺失,1例为AZFb、AZFc区共同缺失。结论:①精液异常VC不育与Y染色体微缺失有关;②VC不育患者特别是无精子症和严重少精子症患者,应该进行Y染色体微缺失的检测。     

精子生成障碍患者Y染色体AZFc区部分缺失分析

目的:探讨人类Y染色体无精子症因子C区(AZFc)部分缺失对男性精子生成的影响。方法:选择Y染色体AZFc区9个序列标签位点(STS)sY1258、sY1291、sY254、sY255、sY1201、sY1206、sY1161、sY1197、sY1191,以ZFX/ZFY(X/Y连锁锌指蛋白基因)和SRY(sY14)基因为内对照。对160例Y染色体微缺失筛查均未发现缺失的无精子症及严重少精子症患者,76例正常生育男性DNA进行多重PCR扩增。疑有DAZ基因缺失的个体,采用基因单核苷酸变异分析(single nucleotide variants,SNV)技术,对DAZ基因4个拷贝中的单核苷酸多态位点进行检测,以确定DAZ基因的拷贝缺失类型。结果:160例无精子症及严重少精子症患者(病例组)gr/gr(sY1291)缺失10例,占6.3%;b2/b3(sY1191)缺失14例,占8.8%;新发现sY1291,sY1197缺失1例,占0.6%;b1/b2缺失1例,占0.6%;b1/b3缺失1例,占0.6%。76例正常生育男性(对照组)检出gr/gr缺失4例,占5.3%;b2/b3缺失4例,占5.3%。gr/gr缺失和b1/b3缺失(对照组和病例组)SNV分析均为DAZ1/DAZ2缺失;b2/b3缺失(对照组和病例组)SNV分析均为DAZ3/DAZ4缺失。1例sY1291,sY1197缺失的DAZ-SNVsY587位点缺失,1例b1/b2缺失者DAZ基因未缺失。结论:b2/b3(sY1191)缺失、gr/gr(sY1291)缺失在我国正常人群中多见,为基因组多态性;b1/b2缺失、b1/b3缺失和sY1291,sY1197缺失可能是导致精子生成障碍的高风险因子。     

原因不明性无精症和少精症Y染色体微缺失的筛查分析

目的 探讨原因不明性无精症和少精症不男性与Y染色体微缺失的关系。方法 应用多重PCR技术,对38例原因不明性无精症和少精症者（无精症11例、严重少精症9例、少精症18例）基因组DNA进行Y染色体连锁的18个序列标记位点缺失检测。结果 38例中发现Y染色体微缺失6例,缺失率为16％,其中无精症2例,严重少精症1例,少精症3例。缺失形式前两者为AZFd(DYS 237) AZFc(DAZ DYS240),后者为AZFd(DYS237)。结论 Y染色体微缺失是原因不明性无精症和少精症的重要原因之一。采用多重PCR技术进行缺失检测,是一种非常有效的方法。     

严重少精子症及非梗阻性无精子症患者Y染色体微缺失分析

目的探讨严重少精子症及非梗阻性无精子症与Y染色体长臂微缺失之间的关系。方法该病例对照研究包括216例严重少精子症、189例非梗阻性无精子症患者及100例精液参数正常的对照。采用多重PCR对Y染色体AZFa、AZFb、AZFc及AZFd区域进行检测。玷果在严重性少精子症患者中,AZF总缺失率为10．65％（23／216）,其中以AZFc区缺失最常见,占缺失的78．26％（18／23）;在非梗阻性无精子症患者中,AZF总缺失率为13．76％（26／189）,其中也以AZFc区缺失最常见,占缺失的57．69％（15／26）;在正常对照中发现1例AZFb缺失,两病例组AZF区缺失分别与对照组相比较均具有显著差异（X^2=9．066,P=0．003;X^2=10．74,P=0．001）。结论通过对Y染色体微缺失的检查可以从基因水平寻找生精障碍的原因以及为优生优育提供可靠的遗传信息依据。     

Molecular detection of Y chromosome microdeletions: an Irish study

The region of the Y chromosome most critical for male fertility is called the azoospermia factor (AZF) region and it is located within subintervals five and six on the long arm of the Y chromosome. Several genes, all residing here, contribute to spermatogenesis and deletions in these genes are thought to be pathogenetically involved in some cases of male infertility associated with azoospermia or oligozoospermia. The aim of this study was to establish the prevalence of microdeletions in the AZF region of the Y chromosome in an Irish male population undergoing fertility treatment. To do this, we applied and compared two independent polymerase chain reaction (PCR) based screening methods, namely, a PCR protocol using several sequence-tagged site (STS) primer sets and a recently published multiplex PCR Y chromosome screening protocol. A total of 78 patients, attending the IVF unit at University College Hospital, Galway, were included in this study. Of them, 56 suffered from idiopathic azoospermic/oligozoospermic infertility. The remaining 22 patients had various conditions, which may have contributed to their infertility. A total of 50 age-matched normospermic men were included as controls. Two microdeletions were found; one in the AZFa region and one in AZFb region. These deletions were observed among the truly idiopathic cases. Further analysis was performed to study the extent of the deletions and it was confirmed that each deletion encompassed the respective AZF region including the AZF candidate gene.     

PCR analysis of Yq microdeletions in infertile males,a study from South India

Aim: To estimate the frequency of microdeletions in the long arm of Y-chromosome of 20 infertile males from South India. Methods: Polymerase chain reaction (PCR) amplification using Y-specific STS of azoospermJa factor (AZF) regions i.e., SY 84 for AZFa, SY 127 for AZFb and SY 254 for AZFc. Results: Of the 20 infertile subjects 3 (15%), one azoospermic and two oligozoospermic, showed microdeletions in the AZF region of Ychromosome. Conclusion: The frequency of deletions involving AZF region of the Y-chromosome is 15% in azoospermic and severely oligozoospermic infertile men. PCR amplification of AZF locus is useful for the diagnosis of microdeletions in the Y-chromosome. (Asian J Androl 2002 Dec; 4: 265-268)     

Near demise of a child with Prader-Willi syndrome during elective orchidopexy

The case of a morbidly obese 3.5-year-old boy, with Prader-Willi syndrome (PWS), who experienced a life-threatening episode of pulmonary edema soon after induction of general anesthesia with sevoflurane and intubation for orchidopexy is presented. The patient who had history of sleep apnea and who had an uneventful laparoscopy under general anesthesia 6 months previously was supported with mechanical ventilation with positive end expiratory pressure but developed hyperthermia, pneumonia, sepsis, and Acute Respiratory Distress Syndrome in the intensive care unit. He recovered fully 11 days after surgery. The possible contributing factors for the development of pulmonary edema are discussed. Arrangements for monitoring in an intensive care setting after surgery are highly recommended for patients with PWS.     

Microdissection TESE is superior to conventional TESE in patients with nonobstructive azoospermia caused by Y chromosome microdeletions

Nonobstructive azoospermia is caused in up to 10% by microdeletions of the Y chromosome in the azoospermia factor (AZF) region, which is divided into three nonoverlapping areas (AZFa, AZFb and AZFc). In 25 male patients with AZF microdeletions, the results of two different techniques for surgical sperm retrieval (SR), conventional multilocular TESE and microdissection TESE, were studied retrospectively over a period of 19 years. Conventional multilocular TESE was carried out in 11 patients and microdissection TESE in 14 patients. Successful SR was possible only in patients with isolated AZFc microdeletions, so only the 20 patients with AZFc microdeletions alone were taken into account for the comparison of the both operative techniques. The sperm detection rate for conventional multilocular TESE was 25%, the sperm detection for microdissection TESE was significantly higher with 67%. In all patients, a histological examination of the testicular tissue was carried out, which showed a mixed picture, but Sertoli‐cell‐only syndrome in most cases. FSH was no prognostic marker for successful SR. In two of six couples performing an intracytoplasmic sperm injection until now, a pregnancy occurred.     

Y染色体新缺失类型gr/gr微缺失对精子发生的影响

目的探讨Y染色体新缺失类型gr/gr微缺失对睾丸精子发生的影响。方法选取Y染色体特异性序列标签点(STS),用多重PCR技术检测严重男性不育患者118例,志愿捐精者120例,观察Y染色体gr/gr微缺失情况。不育患者经2次以上精液分析,均为睾丸性无精子症或精子密度<1×106/ml。捐精者均经精液分析和染色体检查,符合卫生部标准的正常男性。追踪分析6例gr/gr微缺失者父亲微缺失情况,其中3例为不育者的父亲,另3例为捐精者的父亲。结果多重PCR检测118例男性不育患者gr/gr微缺失12例(10.2%);120例捐精者gr/gr微缺失11例(9.2%)。2组比较差异无统计学意义(P>0.05)。6例gr/gr微缺失者的父亲均表现gr/gr微缺失。结论Y染色体gr/gr微缺失既存在于严重不育男性,也存在于生精功能正常的捐精者中,gr/gr微缺失可能遗传自父亲,可能未影响精子的发生,不能认为是精子发生的遗传风险因子。     

中国男性不育患者染色体核型及Y染色体微缺失分析

目的 探讨男性不育症与染色体畸变及Y染色体微缺失之间的关系.方法临床诊断男性不育患者1975例,采集外周血淋巴细胞常规培养,Giemsa染色,镜下观察并分析染色体核型;选取Y染色体特异性序列标签点(STS),应用PCR技术对无精子症及少精子症患者进行Y染色体微缺失检测.结果 1975例患者中,染色体核型异常305例(15.44%),其中常染色体异常101例(5.11%),患者主要表现为少精子症、畸形精子症;性染色体异常204例(10.33%),主要表现以克氏征(5.62%)为主.728例无精子症或少精子症患者中,Y染色体微缺失109例(14.97%),其中AZFa区缺失3例(2.75%),均表现为无精子症;AZFb区缺失5例(4.59%),表现为无精子症2例、严重少精子症2例,精液正常1例;AZFc区缺失者68例(62.39%),患者主要表现为无精子症和严重少精子症;AZFa区和AZFc区均缺失者5例(4.59%),均表现为无精子症;AZFb区和AZFc区均缺失者15例(13.76%),患者以无精子症表现为主;AZFa区、AZFb区和AZFc区均缺失者6例(5.50%),均表现为无精子症.结论染色体异常及Y染色体微缺失均为男性不育的重要病因.

Abstract：

Objective To study the relationship between chromosomal abnormality and Y chromosome microdeletions and male infertility. Methods Lymphocytes were cultured from peripheral blood of 1975 male infertility patients and stained with Giemsa. The chromosomes were analyzed under microscope. Y chromosome specific sequence tags (STS) were selected, then the Y chromosome microdeletions in AZF regions were screened by polymerase chain reaction (PCR) in azoospermia and oligozoospermia patients. Results There were 305 cases of detected chromosomal abnormalities (15.44%) in the 1975 cases. There were 101 cases (5.11 %) with autosome abnormalities which clinically manifested as oligozoospermia and teratospermia. There were 204 cases (10. 33%) of sexual chromosome abnormalities and the patients were mainly characterized with Klinefelter's syndrome. Y chromosome microdeletions were detected in 109 (14.97 %) of the 728 cases of azoospermia or oligozoospermia. The most common microdeletion of Y chromosome was AZFc (62.39%) and these patients were characterized with azoospermia and oligozoospermia. Five patients (4. 59%) who suffered Y chromosome microdeletion in AZFa region and AZFb region were characterized with azoospermia. Fifteen cases (13.76%) with microdeletion in AZFb region and AZFc region were mainly characterized with azoospermia. There were 6 cases (5. 50 % ) of microdeletion in AZFa, AZFb and AZFc regions,these patients were all characterized with azoospermia. Conclusions Both Chromosome abnormalities and Y chromosome microdeletions are important causes for male infertility.