不育症精液生精细胞凋亡率检测

目的了解精液中生精细胞的状况和生精细胞凋亡率与不育症的关系,并作为评估睾丸功能的指标。方法293例精液标本用瑞-姬染色,油镜下进行生精细胞形态学及凋亡率检出与分析。并采用DNA缺口原位末端标记技术(TUNEL)染色进行对照,确认瑞-姬染色对生精细胞凋亡鉴别的可行性。结果在293例精液中,有生精细胞者273例占93.1%;无生精细胞者20例占6.9%。生精细胞凋亡率随精子数量减少而增加,以无精子症者最高,比生育组高5.8倍(P<0.001)。显示瑞-姬染色的可行性与优越性。结论睾丸生精障碍与生精细胞过度凋亡密切相关,可在精液中检出脱落生精细胞,根据精液生精细胞凋亡率可作为评估睾丸功能的指标之一。     

获得性免疫缺陷综合征合并败血症患者的病原谱分析

目的分析本地区获得性免疫缺陷综合征(AIDS)合并败血症患者的病原谱分布以及与CD4~+T淋巴细胞水平的相关性。方法回顾性分析2014年1月至2015年12月武汉大学中南医院收治的AIDS合并败血症患者的住院病历资料,总结病原微生物分布比例,分析病原微生物谱与CD4~+T细胞的相关性。结果 84例AIDS合并败血症患者中,培养结果为细菌感染者38例(占45.3%),真菌感染者39例(占46.4%),混合感染者7例(占8.3%);其中混合感染者均为细菌和真菌二重感染。标本中共分离出91株微生物,最常见的微生物分别为马尼尔菲青霉菌24株(占26.4%)、新型隐球菌21株(占23.1%)、人葡萄球菌9株(占9.9%)和屎肠球菌7株(占7.7%)。83例成人患者中,CD4~+T细胞100个/μl者14例(占16.9%),均为细菌感染;51~100个/μl者5例(占6.0%);CD4~+T细胞≤50个/μl者64例(占77.1%),以真菌和混合感染为主(占67.2%)。CD4~+T细胞≤50个/μl的AIDS合并败血症患者真菌和混合感染败血症的风险显著高于CD4~+T细胞50个/μl者,差异具有统计学意义(OR=6.63;χ~2=15.67,P0.001)。结论 CD4~+T细胞≤50个/μl的AIDS患者为真菌败血症和混合感染败血症的高危人群,本地区应重视AIDS合并马尼尔菲青霉菌和新型隐球菌败血症,须高度重视并注意消毒隔离,避免住院AIDS患者发生院内感染。     

精液脱落细胞检查在少精子不育症治疗中的应用

按照黄守烽主编《实用精液细胞学彩色图谱》中精液脱落细胞染色方法，对69例少精子症病人精液脱落细胞进行检查，能较准确地区分精液中的生精细胞、中性粒细胞。并对每高倍镜视野（40X）生精细胞大于5个或中性粒细胞大于5个的少精子症病人分别进行对症治疗，取得了较好的疗效。提示精液脱落细胞染色检查在少精子不育症的诊断治疗中有较重要的意义。精液脱落细胞检查在少精子不育症治疗中的应用@敖小龙$江西省上高县妇幼保健所!宜春上高县，336400精液;;脱落细胞;;不育症     

精子凋亡和生精细胞凋亡检测与精索静脉曲张的关系

精索静脉曲张(varicocele,VC)常人发病率在8%～20%,在男性不育症患者可达39%.协和医院统计1310例男性不育症中VC为466例,占35.57%[1].柯明辉(2010)统计中日友好医院不育症2599例,有VC者703例,占27.05%.VC是慢性渐进性对睾丸生殖功能有损伤的疾病,每增长1岁精液异常危险性提高0.023倍(2.3/100倍),是非VC精液异常危险率的1.2倍[2].发生于左侧睾丸者约为70%～90%,虽然发生在单侧,但它是一种双侧睾丸损伤性疾病,还具有累积性损害的特征,长时间的VC会引起睾丸功能下降,出现睾丸生殖功能障碍[3].青少年患者应该尽早治疗[4].本文就VC精子形态学、精子和生精细胞凋亡检出率与VC的关系综述如下.     

精液生精细胞检查与男科疾病

目的：探讨精液生精细胞检查与疾病的关系。方法：采用瑞-姬染色法观察精液生精细胞形态、数量及存在比例。结果：化疗、放疗病人出现生精细胞形态异常；长期服用雷公藤药物生精停滞于初级精母细胞阶段；长期接触高温、肾功能不全而行血透者生精细胞停滞于初级精母细胞及精子细胞阶段；双侧隐睾及Klinefelter综合征病人精液中无精子及生精细胞；阴囊鞘膜积液约20％的病人生精停滞于精子细胞阶段；精索静脉曲张（Ⅱ～Ⅲ°）精液中以精子细胞为主；睾丸及附睾炎症病人精液中除出现次级精母细胞、精子细胞外，以中性粒细胞、淋巴细胞、单核巨噬细胞多见。结论：本研究为精液常规检查增加了新内容，为诊断与治疗不育症及环境遗传因索引起的生精障碍，提供了客观指标。     

精液液化异常者精浆中酸性磷酸酶和锌的检测及其临床意义

精液液化异常是指射精后1h未液化的现象,它包括精液不液化和液化迟缓。精液不液化或液化迟缓均可导致不育。Nilson报告在精子数大于20×106/ml病人中,以精液不液化为唯一异常指标者占11.8%;国内胡毓安等在对1000例不育男性精液常规的调查中发现14.8%的病人精液液化不良[1]。多年来的研究发现,精液中的液化因子主要来源于前列腺,精液液化主要受前列腺分泌功能状态影响。为此,我们测定了273例精浆中锌和酸性磷酸酶含量并作白细胞染色计数,以观察它们对精液液化异常的影响。材料和方法1.检测对象:本组273例,其中生育组22例,年龄23￣38岁,被检…     

前列腺液白细胞计数与细菌培养的关系

目的 探讨前列腺液白细胞计数与细菌培养的关系。方法 对2002年9月至2005年1月间，我院泌尿外科门诊962例前列腺炎患者行前列腺液常规检查，计数白细胞。白细胞数目〉10个／高倍镜者为观察组共385例，白细胞数目〈10个／高倍镜者为对照组共577例，同时行前列腺液需氧细菌培养以及支原体、衣原体培养。结果 962例前列腺液标本中，有244例（25．4％）病原体培养阳性，共计培养出病原体285株。研究组白细胞数目为＋、＋＋、＋＋＋、＋＋＋＋时．细菌培养阳性率分别为19．6％、22．5％、31．0％、47．0％、89．3％，对照组前列腺液细菌培养阳性率为19．6％，两组间差别有显著性意义（P〈0．05）。结论 随着前列腺液中自细胞数目的增加，细菌培养阳性率逐渐增高，有助于早期预测细菌培养结果。     

慢性前列腺炎/盆腔疼痛综合征不同于非细菌性前列腺炎和前列腺痛

新的分类方法中 ,慢性前列腺炎 /盆腔疼痛综合征 (ＣＰＰＳ)的定义基于前列腺液、前列腺按摩后尿或精液分析中有或无白细胞。作者将此种分类方法中上述 3个样本的评价与传统分类方法中单独应用前列腺液作为指标进行了比较这是一个前瞻性的临床和实验室研究方案 ,用于评价有症状但无尿道炎、急性细菌性前列腺炎或慢性细菌性前列腺炎患者。结果 :临床和微生物学评价了 310例临床诊断为前列腺炎患者 ,选择 140例有前列腺液、前列腺按摩后尿、精液标本者 ,共 42 0个标本。 111/ 42 0 (2 6 % )个标本证实有炎症 ,包括 39个前列腺液标本 (白细胞≥…     

男性生殖道非特异性感染性不育的精液变化

本文对36例生殖道非特异性感染性不育者的精液进行了研究分析,同时选择11例正常生育力者的精液作为对照组。研究分析的项目包括:精液或前列腺液的微生物学检查、精液质量分析和精浆免疫球蛋白IgG、IsA测定。经测定的结果表明,生殖道非特异性感染对精液质量具有不同程度的影响。当精液或前列腺液中的菌落计数>5千或白细胞>20只/HP时,精液的粘稠度、精子自身凝集、精子畸形率以及精浆中免疫球蛋白IgG、IgA都有明显增加而精子活率、活动力和精子平均运动速度均有不同程度的下降。     

精液白细胞含量变化对精子顶体酶活性的影响

目的观察精液白细胞含量变化对精子顶体酶活性的影响。方法在抗炎治疗前后采用正甲苯胺蓝过氧化物酶法对精液中白细胞进行定量测定,并与精子顶体酶活性进行相关性分析。结果383例男性不育就诊者中有106例精液白细胞＞1×10~6个/ml,其精子项体酶活性(12.81±9.23)μIU/10~6精子；经抗炎治疗后75例精液白细胞≤1×10~6个/ml,设为有效组,其精子顶体酶活性(20.65±10.05)μIU/10~6精子,余31例精液白细胞＞1×10~6个/ml,设为无效组,其精子顶体酶活性(13.26±9.81)μIU/10~6精子。治疗后两组精子顶体酶活性有显著性差异(P＜0.01)。结论精液白细胞含量变化可影响精子顶体酶活性,精液白细胞症经抗炎治疗改善后可提高精子顶体酶活性。     

Chlamydia trachomatis infection in male infertility--the clinical usefulness of the detection of antibodies against Chlamydia trachomatis]

BACKGROUND: The objective of this study is to investigate the clinical usefulness of the detection of antibodies against Chlamydia trachomatis (C. trachomatis) in potential male infertility patients. PATIENTS AND METHODS: Analysis of serum samples for the presence of antibodies against C. trachomatis by enzyme immunoassay was performed in 83 male infertility patients. RESULTS: Immunoglobulin A (IgA) and G (IgG)-antibodies against C. trachomatis were found in 16 and 10 of 83 serum samples (19.3 and 12.0%) respectively. IgA and/or IgG-antibodies against C. trachomatis were found in 20 of 83 serum samples (24.1%). Serum IgA, IgG, or IgA and/or IgG positive patients showed increased semen and urine leukocytes numbers and reduced semen volume and sperm numbers, compared with negative patients, respectively, although these effects are not statistically significant. By Spearman's correlation analysis, however, significant positive correlations with serum IgA or IgG antibody index were found in semen and urine leukocytes numbers. Significant negative correlations with serum IgA antibody index was found in semen volume. In serum IgA positive patients, only 1 of 8 first-voided urine samples was positive for C. trachomatis-DNA by PCR. Moreover, the positive rates of surum IgA antibody and cervical antigen in the 8 female partners of IgA positive patients were only 25% (2 of 8 serum samples) and 12.5% (1 of 8 cervical specimens) respectively. CONCLUSION: There was a high prevalence (24.1%) of serum IgA and/or IgG-antibodies against C. trachomatis from infertility patients. The existance of antibodies against C. trachomatis in serum samples possibly influence semen quality, suggesting that C. trachomatis infection may play an important role in male infertility.     

人精液中未成熟生殖细胞的检测及临床意义

目的 :探讨检测人精液中未成熟生殖细胞含量的临床意义。　方法 :运用非连续梯度Percoll法分离人精液中的各类细胞 ,May Grunwald Giemsa染色及FITC标记的CD4 5单克隆抗体免疫荧光鉴定未成熟生殖细胞及白细胞 ,并检测 30份精液中未成熟生殖细胞含量 ,其中生育组 10例 ,不育组 2 0例。　结果 :精液中未成熟生殖细胞主要集中分布于 30 %～ 4 5 %Percoll分离液中 ,白细胞则分布在 5 0 %～ 5 5 %的分离液中。未成熟生殖细胞在生育组为(0 .70± 0 .4 0 )× 10 6/ml,不育组为 (1.2 8± 0 .70 )× 10 6/ml,两组差异有显著性 (P <0 .0 5 ) ,与精子密度、活率及正常形态百分率无明显相关性 (密度r=0 .0 0 2 ,P >0 .0 5 ;活率r=- 0 .2 2 7,P >0 .0 5 ;正常形态百分率r=0 .2 4 0 ,P >0 .0 5 )。　结论 :运用非连续梯度Percoll法能有效地分离精液中未成熟的生殖细胞 ,其含量有可能作为一项临床指标反映精液质量。     

Development of normal reference values for seminal reactive oxygen species and their correlation with leukocytes and semen parameters in a fertile population

Although reactive oxygen species (ROSs) are clearly implicated in the pathogenesis of male infertility, few studies have attempted to define the basal levels of ROSs in fertile men. Levels of ROSs are highly influenced by the presence of leukocytes and are associated with decreased seminal parameters. The objective of our study was to determine the normal ROS reference values in neat and washed semen of a fertile population and to correlate the leukocyte concentrations with seminal parameters. We evaluated 114 fertile men seeking vasectomy and 47 subfertile patients as a positive control. All samples were subjected to semen analysis and Endtz testing; chemiluminescence assay was used to determine ROS levels. All seminal parameters were significantly higher in the fertile men than in the subfertile patients. In nonleukocytospermic samples, ROS levels were lower in the fertile men than in the subfertile patients in neat (0.29 [0.18, 0.54] vs 0.94 [0.38, 1.51]) (P = .001) and washed semen (5.73 [1.90, 14.71] vs 23.4 [9.46, 115.55]) (P = .001). Similarly, in samples with leukocytes (Entdz, less than 1 x 10(6)/mL), ROS levels were lower in the fertile men in neat (0.75 [0.27, 1.71] vs 2.0 [0.97, 27.41]) (P = .001) and washed semen (15.85 [4.18, 62.16] vs 239.83 [33.4, 1193.75]) (P < .0001). As expected, samples with leukocytes had significantly higher ROS values in washed and neat semen. In the fertile population, ROSs were positively correlated with leukocytes and negatively correlated with sperm count and motility. In semen samples without leukocytes, the normality cutoff of ROSs was 0.55 x 10(4) counted photons per minute with 76.4% area under the curve (AUC) in the neat samples and 10.0 x 10(4) counted photons per minute with 77% AUC in the washed samples. In semen samples with leukocytes, the cutoff for ROSs in neat samples was 1.25 with 72.7% AUC and 51.5 with 81% AUC in the washed samples. We defined the cutoff levels of ROSs in a fertile population. Seminal leukocyte levels below 1 x 10(6)/mL were associated with increased ROSs. ROS levels were positively correlated with leukocytes and negatively correlated with sperm motility and concentration. Patients with normal seminal parameters and lower seminal leukocyte levels may benefit from therapeutic interventions that improve semen quality.     

Reactive oxygen species generation by seminal cells during cryopreservation

Objectives. To test the hypothesis that conventionally used procedures for semen cryopreservation may cause an increase in the production of reactive oxygen species (ROS) by sperm or by seminal leukocytes, which may contribute to poor sperm function following cryopreservation.Methods. Eighteen semen specimens with normal parameters from healthy male donors 22 to 40 years of age were each divided into two portions. The first portion was combined 1:1 with Test Yolk Buffer-Glycerol Freezing Medium and was frozen by gradual cooling into liquid nitrogen (−196°C). The second portion was washed and the cells were resuspended in Sperm Washing Medium (SWM) and incubated at room temperature to serve as controls. After a period of treatment, frozen samples were thawed and semen cells were washed and resuspended in SWM. ROS generation by semen cells from each treatment group was measured on a luminometer. Sperm motility, sperm viability, and sperm membrane integrity were also measured in both control and freeze-thaw samples. To further assess ROS generation by semen cells during the cooling process, aliquots of washed semen cells and purified polymorphonuclear leukocytes (PMNs) were incubated separately at different temperature conditions (37°C, 22°C, 4°C, and −20°C). ROS activity in each treatment group was measured and compared with each other.Results. In both semen cells and PMNs, ROS activity increased significantly during the cooling process. The highest ROS levels were recorded in both groups when cooled to 4°C. The ROS levels were extremely low in samples cooled to −20°C and in freeze-thaw samples, probably due to marked loss of cell viability.Conclusions. Gradual reduction of temperature during the process of semen cryopreservation can cause a significant ROS generation by semen cells. ROS is particularly elevated during cooling if the semen sample is contaminated by more than 0.5 × 10⁶ leukocytes. Removal of leukocytes from semen samples or treatment with antioxidants prior to cryopreservation may improve sperm viability and function.     

Relationship between seminal white blood cell counts and oxidative stress in men treated at an infertility clinic.

In semen, granulocytes are major producers of reactive oxygen species (ROS), which can damage sperm. The diagnosis of leukocytospermia is usually based on the World Health Organization (WHO) definition of 1 x 10(6) white blood cells per milliliter, but controversy remains over the minimum leukocyte level that impairs fertility. The goals of this study were to clarity the relationship between leukocyte count and oxidative stress and to establish the minimum leukocyte count associated with oxidative stress. To do so, we compared oxidative stress in semen samples with different leukocyte counts (by the Endtz test) after a simple wash-and-resuspend procedure and determined the correlation between leukocyte counts and oxidative stress (expressed as ROS-TAC score, a composite score calculated from ROS levels and total antioxidant capacity (TAC), both measured with chemiluminescence assays). ROS-TAC decreases as oxidative stress rises. We compared specimens from 271 men attending an infertility clinic and 28 healthy controls. About 9% of patients had WHO-defined leukocytospermia and an additional 16% had some leukocytes. Samples with no seminal leukocytes had significantly lower ROS levels and significantly higher ROS-TAC scores than samples with any seminal leukocytes, even very low levels. Oxidative stress was correlated with rising white blood cell (WBC) count (r = .39; P < .001). Receiver operating characteristics curves showed that ROS-TAC score would be fairly accurate at distinguishing between patients with any leukocytes and those with no leukocytes (area under the curve, 75%). In conclusion, oxidative stress occurs even in patients with very low seminal WBC counts (between 0 and 1 x 10(6)/mL) and rises with an increase in WBC count. Therefore, we are unable to determine a safe minimum WBC count; the presence of any WBCs is associated with oxidative stress and may therefore impair fertility. Complete removal of WBCs from semen samples used for assisted reproduction may help reduce oxidative stress.     

白细胞精子症的研究进展

白细胞精子症是男性不育的一个重要原因,目前对精液白细胞的分布、来源和作用尚完全不清楚。精液中增多的白细胞及其产物可对精子功能和精液质量产生影响,其中活性氧(ROS)的增加与男性不育关系密切。白细胞精子症的治疗和预防对改善精液质量有一定作用。     

Progress in leukocytospermia research]

Leukocytospermia is a most common cause of male infertility, but the distribution, origin and role of leukocytes in semen are still controversial. Some reports on leukocytospermia have indicated its negative effects on semen parameters and even in vitro fertilization (IVF). Recent literature has made it clear that the most deleterious effect of leukocytospermia is that the increased reactive oxygen species (ROS) may cause sperm damage, leading to significantly increased male infertility. The treatment and prevention of leukocytospermia have been proven of help for improving semen parameters.     

男性不育患者精液阴道加德纳菌感染调查

目的 :了解男性不育患者精液阴道加德纳菌 (Gv)的感染状况。　方法 :收集 2 0 0 2年 4月～ 2 0 0 3年 5月期间无锡市妇幼保健院男性不育门诊就诊的 373例男性精液标本 ,以及其中 6 3例Gv阳性患者的配偶阴道拭子标本 ,应用套式聚合酶链反应 (nPCR)技术进行Gv检测。　结果 :男性不育患者精液Gv定植率为 4 4 .2 % ,阳性者配偶阳性率达 87.3%。　结论 :男性不育患者精液Gv定植率较高 ,Gv可经性生活传播。     

苏州地区2640例男性不育症患者精液质量分析

目的:研究苏州地区男性不育症患者精液质量状况。方法:采用计算机辅助精液分析技术,检测2 640例以不育为主诉的男性精液标本的质量。结果:2 640份精液标本中,各项指标均正常的精液占27.35%;a+b级精子百分率异常者占47.35%(1 250/2 640);精子活动率异常者占42.39%(1 119/2 640)。精子活动率和a+b级精子百分率随患者年龄增加呈降低趋势,尤以40岁以上男性降低更为明显。弱精子症和少精子症发病率分别为37.31%(985/2 640)和8.94%(236/2 640),且30岁以上不育男性随年龄增加有增高趋势。随着精子活动率和精子浓度的下降,精子的9项运动参数VCL、VSL、VAP、MAD、LIN、STR、WOB、ALH、BCF明显降低。结论:苏州地区不育男性主要表现为精子活动率降低,尤其是a+b级精子百分率的降低。由于精子活动率、活力以及少精子症和弱精子症的发病率均与年龄有关,提示男性亦有合适的生育年龄。