非梗阻性无精子症和隐匿精子症与睾丸体积、血FSH和AZF基因微缺失的相关性分析

目的：探讨非梗阻性无精子症和隐匿精子症与睾丸体积、血FSH和AZF基因微缺失的相关性。方法：161例男性不育患者分为非梗阻性无精子症（A组,n=86）、隐匿精子症（B组,n=49）、严重少精子症（c组,n=13）和其它患者（D组,n=13,包括死精子症4例,梗阻性无精子症3例,正常精子1例,其他少弱精子症5例）,进行睾丸体积、血FSH水平和彳砑基因微缺失检测,AZF基因微缺失检测位点包括AZFa（sY84、sY86）、AZFb（sYl27、sYl34）、AZFcd（sY254、sY255、sYl57、sYl45和sYl52）。结果：4组睾丸体积〈12ml的比例分别为73．26％（63／86）、34．69％（17／49）、7．69％（1／13）、30．77％（4／13）;4组FSH水平升高1倍以上的比例分别为67．44％（58／86）、32．65％（16／49）、15．38％（2／13）和O．OO％（o／13）,组间均有极显著统计学差异（P〈O．001）;4组4ZF基因微缺失率分别为15．12％（13／86）．18．37％（9／49）、0．oo％（o／13）和0．00％（0／13）,组间均无统计学差异（P〉0．05）,其中A组爿zn基因缺失1例,AZF（b＋c＋d）缺失5例,AZF（c＋d）基因缺失7例,B组9例均AZFC和（或MZFd基因缺失,c组和D组均未见AZF基因缺失。结论：随着生精功能障碍程度的加重,血FSH水平升高和睾丸体积缩小趋势明显,而与AZF缺失几率无高度相关性;但在非梗阻性无精子症和隐匿精子症组中AZF基因缺失几率仍然偏高,AZFa和AZFb缺失大多出现在非梗阻性无精子症组中,而隐匿精子症组以AZFc和AZFd缺失为主。     

未明确诊断意义的不典型子宫颈鳞状细胞伴DNA倍体异常的临床意义

目的通过宫颈细胞学和DNA倍体分析,探讨未明确诊断意义的不典型鳞状细胞（ASCUS）伴有DNA倍体异常的临床意义。方法选择2010年7月至2011年7月湖北省宜昌市妇幼保健院在夷陵地区进行宫颈癌筛查的635例农村妇女的宫颈细胞学、病理学及DNA倍体分析资料进行分析。将宫颈脱落细胞制成两张玻片,一张巴氏染色行TBS诊断,另一张Feulgen染色行DNA倍体分析。结果在635例宫颈细胞学标本中,正常细胞学418例,ASCUS 145例,不除外高度鳞状上皮内病变（ASC-H）17例,低度鳞状上皮内病变29例,高度鳞状上皮病变26例,其中伴DNA倍体异常（非整倍体细胞）者所占比例分别为2.6%（11/418）、22.8%（33/145）、76.5%（13/17）、89.7%（26/29）和96.2%（25/26）。在阴道镜下活检的145例ASCUS患者中,伴DNA倍体异常者33例,其中10例（30.3%）为CIN2及以上病变;DNA倍体正常者112例,其中9例（8.0%）为CIN2;两者比较,差异有统计学意义（P〈0.01）。17例ASC-H患者中,13例（76.5%）DNA倍体异常者均检出CIN2及以上病变;4例DNA倍体正常者检出1例CIN2,两者比较,差异有统计学意义（P〈0.01）。结论 ASCUS、ASC-H伴有DNA倍体异常患者检出CIN2及以上病变风险增高。     

非梗阻性无精子症患者睾丸组织CREM表达及意义

目的:探讨非梗阻性无精子症(NOA)患者睾丸组织中CREM表达及意义。方法:选取NOA患者28例和生精正常的梗阻性无精子症(OA)患者19例,收集睾丸活检组织,采用RT-PCR、免疫组化法及Western blot法检测睾丸组织中CREM mRNA及CREM蛋白表达。结果:CREM蛋白主要表达于睾丸组织生精小管的精母细胞、精原细胞和早期精子细胞的细胞核。OA睾丸组织中CREM mRNA含量高于NOA睾丸组织[(0.028±0.022) vs (0.014±0.024)],OA睾丸组织中CREM蛋白表达高于NOA睾丸组织[(2.263±0.371) vs (1.145±0.535)],差异均有统计学意义(P0.001)。结论:睾丸组织中CREM mRNA及蛋白表达下调可能是NOA发病的原因之一。     

精液中生精细胞形态学及精液生物化学在无精子症患者诊断及处理中的应用

目的：评价精液中生精细胞形态学及精液生物化学的研究,是否可以对无精子症患的诊断和处理提供依据。方法：精液中成熟和未成熟的生精细胞由睾心潮生成,70%的甘油磷酰基胆碱（GPC）由附睾合成,果糖大部分或全部来自精囊,酸性磷酸酶（ACP）则来自前列腺。本检测了16例正常志愿,12例输精管切除及186例无精子症患的上述参数并对结果进行分析。结果：在输精管切除和阻塞性无精子症的精液中,均未见到成熟与未成熟的生精细胞,且GPC水平在这两组中也显下降。而在非阻塞性无精子症患中,存在有未成熟生精细胞,但GPC、ACP及果糖的水平正常。基于这些参数而作出的阻塞性和非阻塞性无精子症的诊断,与睾丸活检的发现基本一致。在一些由于精子发生低下或精子发生停滞所致的无精子症患中,SGCM的研究对客观地检测生精组织对特殊治疗的反应很有帮助。结论：生精细胞形态学及精粹生物化学的参数是一些非常有价值的非侵入性标志,可以区分阻塞性和非 阻塞性无精子症,有很高的预示性价值。     

固相透明质酸法优选精子的效果观察

目的：对比研究高通量的固相透明质酸（HA）分离新技术较传统密度梯度法在精子优化分离效果方面的优势性。方法：收集新鲜精液标本31份,每份标本液化后被均分成2部分,一部分精液以固相HA法分离精子,即将精液加入到底部表面包被有HA的培养皿中,洗脱掉未结合的精子,分离、收集与HA结合的精子：另外一部分精液以传统的密度梯度法优化分离精子,观察2种方法提取后精子各项研究指标的差异性,以及相对变化幅度。结果：与密度梯度法组相比,固相HA法组处理后精子的正常形态率、获能2h酪氨酸磷酸4~（TP）、ATP（精子获能2hTP率一未获能精子TP率）、诱发顶体反应（AR）和△AR（诱发AR率一自发AR率）等指标明显增高CP〈0．001）,增长幅度依次为：1．55倍（95％CI=1．04,~2．81）、1．81-fg-~（95％CI=0．89-6．11）、3．35-9}（95％CI=1．04-10．32）、1．37倍（95％CI=0．96～2．71）和1．88倍（95％CI=1．09,-4．71）;而固相HA法组自发AR、核蛋白不成熟度和DNA碎片率显著降低（P〈O．001）,分别是密度梯度法组的68％（95％CI=19％-102％）、47％（95％CI=2~／~103％）和21％（95％CI=0％～70％）;2种方法处理后精子的前向运动率和活动率无统计学差异（P〉0．05）。结论：基于成熟精子头部具有HA受体可与外源性HA特异性结合原理的精子。固相HA分离技术,与传统的密度梯度法相比,分离后的精子具有更优的受精潜能和成熟度,可显著提高分离精子的整体质量,有望进一步改善辅助生殖结局。     

非梗阻性无精子症患者的表皮生长因子及受体表达

目的:探讨血清表皮生长因子(EGF)和睾丸组织表皮生长因子受体(EGF-R)与非梗阻性无精子症睾丸生精功能障碍的关系。方法:采用放免法对20例无精子症患者和20例已育男性志愿者进行血清EGF测定;无精子症患者接受睾丸活检,病理检查按Johnson评分法评价睾丸精子发生及障碍的程度;采用免疫组化SABC法对睾丸3种主要细胞进行EGF-R表达的检测。结果:无精子症患者血清EGF浓度明显低于男性对照者(P<0.05)。睾丸活检生精功能评为8分者14例;3分者2例;6、5、4和 2分者各1例。在20例无精子症睾丸的间质细胞、支持细胞和生精细胞均有EGF-R的表达,间质细胞EGF-R强阳性和阳性表达明显低于支持细胞和生精细胞(P<0.05)。结论:无精子症患者血清EGF浓度下降、间质细胞EGF-R强阳性和阳性表达的减少与睾丸生精功能障碍的程度是相一致的,说明血清EGF及睾丸EGF-R在调节睾丸生精功能方面有重要的作用。     

40例输精管道梗阻性无精子症诊疗策略分析

目的:探讨输精管道梗阻性无精子症(OA)的诊疗策略。方法:40例输精管道OA患者为研究对象。首先行输精管道探查,根据术中探查情况行输精管-输精管显微吻合术(VV)、输精管-附睾显微吻合术(VE)、交叉输精管-输精管吻合术、交叉输精管-附睾管显微吻合术或睾丸取精冻存术。结果:精道探查后,共行吻合手术26例,取精+冻精手术14例。40例患者中,共有3例患者失访(2例吻合患者,1例冻精患者)。随访到的24例吻合术患者术后复通率为58.3%(14/24),自然受孕率为29.1%(7/24)。随访到的13例取精+冻精患者中已有11例行ICSI/冻精助孕。结论:输精管道OA,可根据术前严格评估和术中探查行显微复通手术或取精术;复通术后可获得自然妊娠的机会,交叉显微吻合为输精管道复通的有效途径之一。     

新柏氏TCT在宫颈癌筛查中的应用研究

目的 探讨新柏氏TCT检测技术在妇科门诊宫颈癌筛查中的准确性及应用价值。方法 2005年3月-2006年9月在迁安妇幼保健院妇科门诊行新柏氏TCT细胞学检查的女性患者1522例，对细胞学检查结果为阳性者行阴道镜检查并多点活检对照。细胞学诊断采用TBS分级系统。结果 细胞学检查结果细胞学标本1498例满意，24例基本满意。检出鳞状上皮原位癌1例（0．1％）、浸润癌1例（0．1％）、上皮内高度病变3例（0．2％）、上皮内低度病变5例（0．3％）、未明确意义的非典型腺细胞3例（0．2％），未明确意义的非典型鳞状细胞64例（4．2％）、正常范围内197例（12．9％），良性反应性改变1248例（82．0％）。其中滴虫感染79例，占5．2％；念珠菌感染51例，占3．4％；HPV感染8例，占0．5％。结论 新柏氏TCT细胞学检查适用于临床宫颈癌的筛查。     

定量分析卵巢癌细胞中p~（53）基因蛋白表达的意义

目的研究卵巢癌ｐ（５３）基因蛋白表达的量与ＤＮＡ倍体水平、细胞增殖活性及组织学分级的关系。方法采用流式免疫荧光技术对卵巢癌４２例的常规石蜡标本单细胞悬液行ＤＮＡ含量和ｐ（５３）蛋白单克隆抗体免疫荧光标记定量测定。结果４２倒卵巢癌的异倍体率为７６．２％（３２／４２）。细胞增殖指数（ＰＩ）为３１．２４±５．３６。ｐ（５３）蛋白表达的阳性率为５９．５％（２５／４２），其表达的量与ＤＮＡ倍体水平、组织学分级及ＰＩ均相关（Ｐ＜０．０５，Ｐ＜０．０１），ＤＮＡ异倍体率与组织学分级亦有关（Ｐ＜０．０５）。卵巢癌ｐ（５３）蛋白表达阳性的ＤＮＡ含量、ＰＩ值均显著高于阴性者；组织学分化越差，ｐ（５３）蛋白表达阳性率越高。结论ｐ（５３）蛋白表达阳性可作为反映卵巢癌恶性程度的重要指标。     

液基细胞学、HPV检测及DNA倍体定量分析在宫颈癌筛查中的价值

目的 探究液基细胞学、HPV检测及DNA倍体定量分析在宫颈癌筛查中的价值。方法 择取2019年1月至2022年1月于山东省立第三医院行宫颈癌筛查的1 200例患者进行研究。所有患者均行液基细胞学检测、HPV检测及DNA倍体定量分析,并以宫颈活组织病理检查结果为金标准,比较液基细胞学检测、HPV检测及DNA倍体定量分析三种检测方法的敏感性、特异性及准确性。结果 液基细胞学、HPV检测及DNA倍体定量分析筛检宫颈癌,敏感性及准确性比较,差异均无统计学意义（P>0.05）;特异性比较,液基细胞学检测最高,HPV检测最低,差异有统计学意义（P <0.05）。结论 液基细胞学联合HPV检测及DNA倍体定量分析测能够显著提高宫颈癌筛检准确性,实现对宫颈疾病早发现早治疗,继而降低宫颈癌的发病率及死亡率。     

转录因子Ets差异基因5(ETV5)基因多态性与非梗阻性无精子症相关性研究

目的:探讨转录因子Ets差异基因5(ETV5)基因多态性与非梗阻性无精子症的易感性的相关性。方法:应用Sequenom MassArray质谱阵列技术对368例已生育的汉族男性人群(对照组)和361例汉族男性非梗阻性无精子症(病例组)ETV5基因的5个标签单核苷酸多态(single nucleotide polymorphism,SNP)位点(rs12631658,rs6444106,rs7430047,rs7433760,rs9824882)进行基因型检测。应用Plink1.07软件对数据资料进行统计分析,比较对照组与病例组最小等位基因频率(MAF)及基因型差异,运用Haploview软件对ETV5基因进行单体型分析。结果:ETV5基因5个标签SNP的等位基因频率、基因型分布组间比较均无统计学差异(P0.05),进一步的单体型分析亦未显示有统计学差异(P0.05)。结论:ETV5基因5个标签SNP位点多态性与汉族男性非梗阻性无精子症的发生可能不相关。     

Seminal plasma characteristics as indicators of cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations in men with obstructive azoospermia

Objective: To determine the prognostic value of seminal plasma volume, pH, fructose, and -glucosidase for the detection of cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations.

Design: Retrospective data analysis.

Setting: University infertility clinic (referral center).

Patient(s): Fifty-nine men with obstructive azoospermia.

Intervention(s): Semen analysis including seminal plasma volume, pH, fructose, -glucosidase, molecular genetic diagnosis of CFTR mutations and FSH measurement.

Main Outcome Measure(s): Sensitivity and specificity of seminal plasma markers for the detection of CFTR mutations.

Result(s): A CFTR mutation was detected in 26 of 59 patients with obstructive azoospermia. Patients carrying a mutation had significantly lower seminal plasma volume (mean ± SEM: 1.5 ± 1.4 mL vs. 2.8 ± 2.2 mL), lower pH levels (25th percentile, median, 75th percentile: 6.5, 6.8, 7.5 vs. 7.7, 7.9, 7.9) and lower fructose content (1.0, 1.1, 3.7 vs. 5.8, 20.0, 83.0 μmol/ejaculate) than those without mutations. Diagnostic efficacy for detection of mutations was best (pH 81.4%, fructose 81.8%) at a cutoff level for pH of 7.4 and fructose of 2 μmol/ejaculate.

Conclusion(s): Seminal plasma markers provide an effective, noninvasive method to predict CFTR mutations in men with obstructive azoospermia.     

人子宫内膜细胞体外诱导蜕膜化的效果观察

目的:观察孕激素(progestogen,P)、雌激素(estrogen,E_2)和8-溴-环磷酸腺苷(8-Br-cAMP)不同组合的3种方法对人子宫内膜细胞体外诱导蜕膜化的效果。方法:收集因子宫良性疾病行全子宫切除术患者的子宫内膜,分离纯化子宫内膜间质细胞(endometrial stromal cell,ESC),进行细胞体外培养并传代,免疫荧光鉴定细胞类型。分别采用P+E_2(PE组)、8-Br-cAMP(PC组)、P+E_2+8-BrcAMP(PEC组)3种不同组合方法蜕膜化诱导处理第3代ESC,并以空白对照为对照组。显微镜下观察蜕膜化过程中的细胞形态变化,化学发光法检测3种不同蜕膜化诱导方法处理24h.48h及96h后细胞培养液中催乳素(PRL)水平,比较3种蜕膜化方法的效果。结果:蜕膜化过程中ESC形态从长梭形逐渐变为圆形,细胞体积变大,胞核增大,部分出现双核或多核,细胞边界变模糊。随着培养时间延长,各蜕膜化诱导组培养液中PRL呈不同水平升高。其中PC组PRL水平及蜕膜化细胞形态均较PE组和PEC组升高明显(P0.05)。结论:P+8-Br-cAMP法,较P+E_2、P+E_2+8-BrcAMP法能更有效地诱导子宫ESC体外发生蜕膜化,是高效可靠、更合适的蜕膜化诱导方法。     

Hormonal therapy for non‐obstructive azoospermia: basic and clinical perspectives

Microdissection testicular sperm extraction (micro‐TESE) combined with intracytoplasmic sperm injection is a standard therapeutic option for patients with non‐obstructive azoospermia (NOA). Hormonal treatment has been believed to be ineffective for NOA because of high gonadotropin levels; however, several studies have stimulated spermatogenesis before or after micro‐TESE by using anti‐estrogens, aromatase inhibitors, and gonadotropins. These results remain controversial; however, it is obvious that some of the patients showed a distinct improvement in sperm retrieval by micro‐TESE, and sperm was observed in the ejaculates of a small number of NOA patients. One potential way to improve spermatogenesis is by optimizing the intratesticular testosterone (ITT) levels. ITT has been shown to be increased after hCG‐based hormonal therapy. The androgen receptor that is located on Sertoli cells plays a major role in spermatogenesis, and other hormonal and non‐hormonal factors may also be involved. Before establishing a new hormonal treatment protocol to stimulate spermatogenesis in NOA patients, further basic investigations regarding the pathophysiology of spermatogenic impairment are needed. Gaining a better understanding of this issue will allow us to tailor a specific treatment for each patient.     

5Alpha-reductase type 2 genes in Japanese males do not appear to be associated with cryptorchidism

OBJECTIVE: To evaluate the occurrence of mutations of 5alpha-reductase type 2 gene in patients with cryptorchidism or idiopathic azoospermia. DESIGN: Controlled clinical study. SETTING: Yamagata University Hospital, Yamagata and Tokyo Electric Power Company Hospital, Tokyo, Japan. PATIENT(S): Patients with isolated cryptorchidism (n = 48) or idiopathic azoospermia (n = 33) and a control group of males with proven fertility (n = 51). INTERVENTION(S): Collection and analysis of blood, leukocytic DNA, semen, and testicular biopsy. MAIN OUTCOME MEASURE(S): Semen analysis, screening of the 5alpha-reductase type 2 gene within exon 1 using direct sequencing and exons 2-5 using single strand conformational polymorphism analysis. RESULT(S): No mutations were detected in isolated cryptorchid men, idiopathic azoospermic men, or controls within the 5 exons of the 5alpha-reductase type 2 gene. CONCLUSION(S): 5Alpha-reductase type 2 gene abnormalities do not constitute a major factor in the development of cryptorchidism or idiopathic azoospermia.     

Seminal acid phosphatase from normal, oligospermic, vasectomized, and azoospermic men

The activity of seminal acid phosphatase was determined in both normal and abnormal human semen samples and the enzyme activity was correlated with total protein content of seminal fluid. Fresh human semen samples were collected from 88 subjects, aged 25-40, after 7 days of abstinence, by masturbation. Semen samples were grouped into 4 according to sperm count: 1) count above 40 million/ml and normal morphology; 2) count below 40 million/ml with normal morphology (oligospermic); 3) azoospermic (sperm absent); and 4) vasectomized males. Acid phosphatase was measured colorimetrically, and protein was assayed by the method of Lowry et al. Despite similar ejaculate volumes in both Groups 1 and 2, the percentage motility of sperm from oligospermic men (Group 2) was significantly less (P=.05). The protein content was found to be highest in oligospermic males, but not significantly higher (P.05). The normal, oligospermic, vasectomized, and azoospermic samples showed a decreasing order of acid phosphatase activity, and a significant difference of activity (P.05) was exhibited only when normal and azoospermic samples were compared. An inverse relationship between acid phosphatase activity and total protein content was found. The normal semen samples with high protein concentrations were found to possess low acid phosphatase activity and samples having low protein content showed approximately 3 times higher acid phosphatase activity. Statistical analysis has shown these are highly significant relationships (P.001 in each case).     

Comparison of intracytoplasmic sperm injection with testicular spermatozoa success in infertile men with obstructive and non-obstructive azoospermia; a retrospective analysis

The aim of this study was to compare the outcome of intracytoplasmic sperm injection (ICSI) and embryo transfer between couples with infertility due to male non-obstructive azoospermia (NOA) and obstructive azoospermia (OA). A retrospective analysis of 234 couples with azoospermia who were treated by ICSI and embryo transfer between January 2007 and October 2010 was performed. There were 61 couples in NOA group and 173 couples in OA group. Fertilization rates, pregnancy and clinical pregnancy rates were the main outcome measures. The number of retrieved mature oocytes, injected oocytes, metaphase II (MII) oocytes, two distinct pronuclei oocytes, cleavage embryos and embryos transferred was not significantly different between the groups. The fertilization rate was significantly lower in NOA group when compared to OA group (56.2 vs. 66.7%, respectively; p?=?0.013) and the pregnancy rate was significantly lower in NOA group than OA group (36.1 vs. 50.9%, respectively; p?=?0.046). The clinical pregnancy rates were not statistically different between the patients with NOA and OA azoospermia groups (24.6 vs. 36.4%, respectively; p?=?0.09). This study suggests that ICSI and embryo transfer together with testicular sperm extraction results in statistically significant lower fertilization and pregnancy rates in men with NOA when compared to men with OA.     

新生儿溶血病与孕妇血清中IgG亚型关系的研究

目的：在已知孕妇血清中IgG抗A（B）效价和其新生儿是否发生HDN的基础上,来探讨孕妇血清中抗A（B）IgG亚型与HDN之间的关系。希望所获得的结果能够弥补目前临床常用的检测O型孕妇体内的抗A03）IgG的效价来预报ABO血型不合HDN的方法的不足,并为临床提供更加有效的预报ABO血型不合HDN的方法。方法：收集2007年10月～2009年5月,在梨园医院输血科进行检查的45例夫妇血型不合。型RhD（＋）孕妇及与其相应的新生儿标本。结果：对孕妇血清中抗A（B）IgG效价进行检测后表明：抗A（B）IgG抗体量与溶血程度有一定的相关性,但并非绝对;不能单纯以孕妇血清抗体效价的高低作为判断胎儿病情是否严重的唯一诊断依据。结论：本实验通过ELISA方法检测孕妇和新生儿血清中抗A03）IgG亚型,发现。型孕妇血清中抗A（B）IgG1是引起ABO血型不合HDN的重要因素,因而检测孕妇体内各IgG亚型的含量比检测抗A03）IgC效价更能准确地预报HDN的发生,并可以指导临床采取有效的预防和治疗措施。     

Seminal plasma inhibin-B level is a useful predictor of the success of conventional testicular sperm extraction in patients with non-obstructive azoospermia

AIM: The value of serum inhibin-B as a predictor of the presence of testicular spermatozoa is still controversial. The purpose of this study is to evaluate the predictive value of the seminal plasma inhibin-B level, which might more directly reflect the secretion by Sertoli cells, and to discriminate between successful and failed testicular sperm extraction (TESE) in non-obstructive azoospermia. METHODS: Sixty-two patients with non-obstructive azoospermia were examined at the Department of Obstetrics and Gynecology at Niigata University Hospital, Niigata, Japan. The level of inhibin-B was measured using a two-site enzyme-linked immunoassay. RESULTS: Testicular sperm were successfully retrieved in 17 of 62 patients (27.4%). The serum levels of follicle-stimulating hormone (FSH) were significantly lower and the serum and seminal inhibin-B concentrations were significantly higher in the successful TESE group compared with the failed TESE group. According to the receiver operating characteristics (ROC) curve analysis, the best discriminating seminal plasma inhibin-B level was 27.0 pg/mL (sensitivity 88.2%, specificity 93.3%). The best discriminating serum inhibin-B level was 34.0 pg/mL (sensitivity 70.6%, specificity 95.6%). The area under the ROC curve for seminal plasma inhibin-B was significantly larger than that for FSH and testicular volume. Using multivariate logistic regression analysis, only seminal plasma inhibin-B was an independent predictor of the presence of spermatozoa on TESE. CONCLUSION: Seminal plasma inhibin-B level is a useful predictor of the presence of testicular sperm in men with non-obstructive azoospermia.