精液分析标准化和精液质量评估——WHO《人类精液检查与处理实验室手册》(第5版)出版

<正>精液分析是评估男子生育力的重要方法,也是男科疾病诊断以及疗效观察的试验依据。精液常规分析易受采集方式、射精频度、温度、实验室条件、检验人员的技术熟练程度及主观判断能力等诸多因素的影响,导致分析结果发生偏差,所以精液的采集与     

不育男性精子染色质结构与精液常规参数的关系

目的 研究不育男性精子染色质结构参数与精液常规参数的关系.方法 采集36例男性不育患者和18例健康对照者的精液标本,用TUNEL法检测精子DNA碎片化,CMA3染色法检测精子染色质包装质量,按照世界卫生组织标准(1999)检测精液常规参数.结果 不育组的精子TUNEL阳性率和CMA3阳性率均显著高于健康对照组,差异具有统计学意义(P＜0.05),精液常规参数正常的不育男性精子TUNEL阳性率和CMA3阳性率也显著高于健康对照组,差异具有统计学意义(P＜0.05),精子TUNEL阳性率和CMA3阳性率与精液常规参数之间具有显著的相关性(P＜0.05).结论 不育男性的精子染色质结构检测结果与健康男性存在显著差异,精子染色质结构检测可提供反映男性生育能力的信息.     

Expression and effects of human telomerase RNA in testicular tumor

Human telomerase RNA (hTR) plays an important role in determining repeated telomere sequence and the expression of an antisense telomerase RNA that leads to telomere shortening and cell death.1 Using highly sensitive in situ nucleic acid hybridisation, we investigated the expression of hTR in human testicular tumours and located its cellular expression. Our study may help in elucidating the role of hTR in human testicular tumours, finding a highly sensitive diagnostic method and a target for gene therapy of testicular tumours.     

人类精子核成熟度检测及其临床意义

人类精子的核成熟是精子结构功能成熟的关键环节,对于维持遗传物质的完整性、保证父方遗传信息的正确传递,具有重要的意义。精子核成熟不良,可能对精子功能和体内外受精结局造成不良影响。近年来出现了多种技术检测人类精子核成熟度,如精子核蛋白提取电泳、苯胺蓝染色等。检测精子核成熟度有助于男性不育发病机制的研究,并可能为男性不育的临床诊断和治疗提供新的思路。     

Tumor Necrosis Factor-related Apoptosis Ligand Induces Apoptosis in Prostate Cancer PC-3M Cell Line

Summary： To study the effect of tumor necrosis factor-related apoptosis inducing ligand （TRAIL） on PC-3M cell line, PC-3M cell line was incubated with gradient concentrations of TRAIL for 4-24 h. Annixin-V fluorescence staining and TUNEL method were employed to detect the apoptosis of PC-3M cells. The morphology of apoptotic PC-3M cells was observed by electron microscopy. The relationship between TRAIL concentrations and the percentage of apoptotic cells was evaluated by flow cytometry. The proliferation inhibitory ratio was calculated by using MTT colorimetry. Our results showed that apoptosis of PC-3M cells could be induced by treatment with TRAIL for at most 4 h. The results of flow cytometry and MTT colorimetry demonstrated a time-and concentration-dependent relationship between cell apoptosis rate and TRAIL concentration. It is concluded that apoptosis of PC-3M cells can be induced by TRAIL. Because of the selective killing effect of TRAIL on tumor cells, it may become a potential alternative for the treatment of advanced prostate cancer.     

非嵌合型Klinefelter综合征患者显微切开睾丸取精及配偶临床妊娠的预测因素分析

目的:探讨非嵌合型Klinefelter综合征患者显微切开睾丸取精术(micro-TESE)的成功率、配偶卵细胞胞质内单精子注射(ICSI)结局,以及可能的预测因素。方法:对49例Klinefelter综合征患者行micro-TESE,并根据取精结果比较成功组与未成功组患者的年龄、生殖激素水平和睾丸体积。成功获取精子者,其配偶行ICSI,随访观察其配偶妊娠、生育情况,并根据妊娠结果比较成功组与未成功组患者及其配偶的年龄、生殖激素水平、睾丸体积和成熟卵母细胞数。同时分析患者micro-TESE及配偶临床妊娠成功与否的可能预测因素。结果:49例患者均经染色体核型分析确定为47,XXY非嵌合体,年龄(28.20±3.52)岁,睾丸体积(2.95±0.84)ml,术前FSH(42.42±14.37) IU/L, LH(22.50±8.64) IU/L,T(6.64±4.13) nmol/L;32例患者成功取到精子,精子获得率(SRR)为65.31%(32/49)。其中29例患者配偶取卵后行ICSI,年龄(26.79±2.97)岁,受精率为(48.14±27.33)%,可利用胚胎率为(63.71±28.90)%,临床妊娠率为48.27%(14/29),已出生7例婴儿,且仍有2例未成功妊娠者剩余冷冻胚胎待移植,7例正在待产。Logistic回归分析显示Klinefelter综合征患者术前T值对其配偶临床妊娠率有预测价值(P=0.015),曲线下面积(AUC)=0.832,cut-off值为5.17 nmol/L。结论:Klinefelter综合征患者接受micro-TESE,将有超过半数者可成功取出精子,配合ICSI技术,部分患者可生育其遗传学后代。同时,患者术前T值5.17 nmol/L提示取精成功后,其配偶有较大可能临床妊娠。     

卵细胞浆内单精子注射在常规治疗失败的死精子症中的应用

目的:评估卵细胞浆内单精子注射(ICSI)在常规治疗失败的死精子症造成的男性不育症中的应用价值。方法:8例经过常规治疗无效的死精子症患者,均表现为精子完全不运动。采用常规ICSI方法治疗,随访受精率、β-HCG阳性率、临床妊娠及分娩等结局参数,并与同期使用活动精子进行ICSI的患者(对照组)对比。结果:死精子症组与对照组在男女方年龄、取卵数上的差异无统计学意义(P>0.05),死精子症组的受精率平均为62.3%(14%～100%),对照组为71.9%(40%～100%),差异无统计学意义(P>0.05)。死精子症组5例患者胚胎移植后血清β-HCG升高,其中4例获得临床妊娠(1例活产,2例继续妊娠,1例孕8+周胚胎停育);对照组5例血清β-HCG升高,均获得临床妊娠,包括3例足月活产和2例继续妊娠。结论:采用ICSI治疗常规治疗失败的死精子症男性不育症患者,受精率及临床妊娠情况令人满意,是一种可行的治疗方法。     

精子染色质结构异常对体外受精-胚胎移植结局的影响

Objective To explore the effects of sperm chromatin structure abnormalities on the outcome of in vitro fertilization and embryo transfer (IVF-ET). Methods Sperm DNA fragmentation and chromatin packaging defects were assessed in 136 couples undergoing IVF-ET because of infertility. The relationship between sperm DNA fragmentation, chromatin packaging defects and fertilization rate and clinical pregnancy was evaluated. Results Both sperm DNA fragmentation and chromatin packaging defect had a negative correlation with fertilization rate (r= -0. 198, P＜0. 05, and r= -0. 389, P＜0. 01,respectively). Both parameters were higher in couples who failed to achieve pregnancy than those who achieved clinical pregnancy (10. 74% vs. 5. 40%, P＜0. 01 and 23. 58% vs. 11. 83%, P＜0. 01,respectively). Conclusion Abnormality of sperm chromatin structure is one of the reasons for IVF-ET failure. Examination of sperm chromatin structure is helpful in predicting he risk of IVF-ET failure and optimizing treatment of infertility.     

非嵌合型克氏综合征显微睾丸取精术后临床妊娠1例报告及文献复习

目的 探讨非嵌合型克氏综合征患者生育遗传学意义后代的可行性.方法 1例就诊于华中科技大学同济医学院附属同济医院的男性患者,年龄29岁,多次精液分析提示无精子,查体双侧睾丸体积小,质硬,约2 ml.性激素水平:卵泡刺激素37.34 IU/L,黄体生成素15.69 IU/L,明显升高;睾酮2.13 ng/ml,稍微偏低.查染色体核型为47,XXY,Y染色体微缺失正常,确诊为非嵌合型克氏综合征.行显微睾丸取精术,同时配合女方取卵后行卵细胞胞质内单精子注射(intracytoplasmic sperm injection, ICSI),随访1个月,观察患者术后并发症及其配偶妊娠情况.结果 显微睾丸取精术中找到少量活动精子,次日女方取卵后行ICSI,3 d后移植2枚新鲜胚胎.术后患者未出现阴囊血肿、感染等并发症,术后随访1个月患者睾酮水平较术前偏低,但性功能未受到影响.女方移植后2周查血HCG,为801 mIU/ml;移植后4周超声提示宫内早孕,单胎,胚胎存活.结论 非嵌合型克氏综合征患者可通过显微睾丸取精术获取精子,同时配合ICSI技术,生育自己遗传学意义的后代.     

肿瘤坏死因子相关凋亡诱导配体诱导前列腺癌细胞凋亡的研究

目的 研究肿瘤坏死因子相关凋亡诱导配体(TRAIL)对前列腺癌细胞的凋亡诱导作用。方法 10μg/L-100μg/L的TRAIL处理培养的PC-3M细胞4～24h后，采用Annexin-V荧光染色、透射电镜、原位末端转移酶标记技术(TUNEL)检测肿瘤细胞凋亡；流式细胞术和MTT比色检测凋亡率、细胞生长抑制率与TRAIL的时间、浓度依赖关系。结果10μg/L～100μg/L TRAIL作用4～24h，肿瘤细胞出现典型的凋亡形态学改变，随着TRAIL浓度的增加或药物作用时间的延长，肿瘤细胞凋亡率也增加，为13．8％～79．6％，同时细胞生长抑制率出现明显增加，药物作用8h为7．5％～37．9％，12h为16．7％～87．9％，24h为39．7％～97．6％。结论 TRAIL能够迅速和显著地诱导PC-3M细胞凋亡，可能成为晚期前列腺癌治疗的新方法。