OBJECTIVE:To compare the computer-assisted sperm analysis (CASA) systems Hamilton-Thorne Integrated Visual Optical System Ⅰ (IVOSⅠ) and IVOS Ⅱ after verifying the performance of the latter so as to ensure the accuracy of the results of analysis.METHODS:Based on the criteria established in the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen (WHO 5th Ed), we compared the main semen parameters obtained from IVOSⅠ with those generated by IVOS Ⅱ, and examined the consistency between the results of the two sperm analyzers.RESULTS:The linear relationship of the outlier test, bias estimation and scatter plot and the results of the outlier test of the two systems all met the requirements of comparison analysis and showed an obvious correlativity. The application scope of the results obtained from the apparatus indicated a reasonable value range, with r = 0.988 for sperm concentration, r = 0.975 for sperm progressive motility (PR), and r = 0.981 for total sperm motility. Evaluation of the acceptability of the predicted bias showed that the allowable total error (TEa) to be 6.67% with sperm concentration at 12 × 106 /ml and 2.34% with PR < 31%, their upper limit of the allowable error < 1/2. The results of IVOS Ⅱ conformed to the requirements of the WHO 5th Ed.CONCLUSIONS:The main parameters derived from IVOSⅠ and IVOS Ⅱ are comparable and consistent, indicating that both can be used for the examination of semen samples.

Objective:To explore the relationship between the level of IL-6,sICAM-1 and immune infertility in male seminal plasma.Methods:189 cases of infertile patients and 100 cases of healthy physical examination people were selected as the research objects from June 2014 to December 2015,and the infertility patients were divided into immune infertility group (88 cases) and non-immune infertility group (101 cases) according to the results of patient's sperm with Mixed Agglutination Reaction (MAR);Patients in immune infertility group were divided into immune positive cells group (WBC is more than or equal to 1 × 106/mL) (36 cases) and immune negative leukocyte negative group (＜ 1 × 106/mL) of 52 cases accordance to the seminal leucocyte peroxidase staining;To analysis and compare the sperm quality,the level of IL-6 and sICAM-1 between the each groups.Results:The rate of sperm forward movement and the survival rate of the sperm in immune infertility group and non-immune infertility group were lower than those of the control group (P＜0.05);There were no differences in semen parameters between the immune infertility group and the non-immune infertility group (P＞0.05);The levels ofIL-6 and sICAM-1 in the immune infertility group were significant higher than those in the non-immune infertility group and the control group (P＜0.05);The levels ofIL-6 and sICAM-1 in the non-immune infertility group were significant higher than those in control group (P＜0.05);Correlation analysis showed that the sperm were IgG adhesion ratio were positively correlated with the level ofIL-6,sICAM-1 (r-=0.438,0.561,P＜0.05) among research object;The levels of IL-6 and sICAM-1 in the patients with immune positive leukocyte were higher than those of the immune negative leukocyte group (P＜0.05).Conclusion:The sperm quality of immune infertility patients were decreased,the higher expression ofIL-6 and sICAM-1 in male semen,the higher MAR positive rate,the greater the possibility of male immune infertility.

Objective:To investigate the relationship between sperm acrosin activity and level of spermatozoa antibody coating in unexplained male infertility.Methods:The sperm acrosin activity of 214 males with unexplained infertility was detected by modified Kennedy method,and the level of sperm membrane antibody by mixed antiglobulin reaction test (MAR).The correlation between sperm acrosin activity and antibody coating of spermatozoa was analyzed.Results:The sperm acrosin activity in 214 unexplained infertile males tested normal in 158 cases (control group) and abnormal in 56 cases (abnormal group).The level of antibody coating of spermatozoa was significantly higher in the abnormal group than that in the control group (P<0.05).There was a negative correlation between sperm acrosin activity and level of spermatozoa antibody coating (r=-0.184,P<0.05).Conclusion:Reduced sperm acrosin activity may be one of the causes for low fertility in males.The presence of antibody coating of spermatozoa may have a certain effect on sperm acrosin activity.

Objective To establish an automatic assay for seminal fructose level and evaluate its accuracy,precision and linear range.Methods The concentration of seminal fructose was measured by hexokinase method on an automatic specific protein analyzer,and the accuracy,precision and linear range of this method were evaluated,and the results of this method were compared with those of indole chromogenic method commonly used in clinical.Results The result of the fructose quality control determined by the automatic method falls within the scope of value.The recovery rates of the recovery experiment were 96.15％ and 103.37％.The automatic method had excellent within-run and between-run precision,and the coefficients of variation (CV) were 0.63％ and 1.27％,respectively.A good linear relationship (r2=1.00) was shown when the concentration of seminal fructose was between 0.065 and 4.16 mmol/L.Compared with the results of indole chromogenic method,the results of automatic method showed statistical difference (P＜0.01) because of different methodologies.But it had a significant positive correlation between the results of seminal fructose concentration from 105 seminal samples analyzed by indole chromogenic and automatic method (r2=0.96,P＜0.01).The linear regression equation:y=1.19x+1.67 (R2=0.93,P＜0.01).Conclusion The established automatic method for analysis of seminal fructose level has good accuracy and precision,and it also has good correlation with the commonly used method in clinical.The operation of this method established in our study is simple,safe and fast.It would be more suitable for analyzing large numbers of samples and it could be applied to andrology laboratory.

Objective:To study the effect of exogenous reactive oxygen species(ROS)contained in cigarette smoke condensate(CSC)on sperm motility in mouse.Methods:Cigarette smoke was dissolved into DMSO to make the cigarette smoke condensate(CSC),which contains high concentration of ROS.Forty five C57BL/6Jmice of 4-week old were randomly divided into three groups:control group,low dose group(0.2mg/ml CSC),and high dose group(2.0mg/ml CSC).The three groups were fed with water with or without different concentration of CSC for 4weeks.After the mice were sacrificed,the CSC toxicity and development of reproductive system were evaluated.In addition,the liver function and sperm morphology,concentration and motility were analyzed.Furthermore,the expression of antioxidant enzymes and pathologic alterations in the testis were examined.Results:There was no significant difference in sperm concentration among the three groups.The sperm motility was significantly decreased in the two treatment groups(19% in the high dose group,32% in low dose group)compared with the controls(61%)(P0.05).The percentage of sperm with rapid progressive motility was 12%,13%,and 24% in the three groups respectively.CSC did not cause toxicity on body development and reproductive system.CSC also did not affect the liver function or the antioxidant gene expression in testis.Conclusions:CSC treatment significantly affects sperm motility,without obvious toxicity on reproductive system,liver function,and body development in mice.This method might be used to establish the asthenozoospermic mouse model.

Objective To investigate the association between the calcium ionophore A23187 induced sperm acrosome reaction rate, total acrosin activity and semen parameters and the IVF outcome, and to compare the predictive value for the male fertility and the outcome of IVF between calcium ionophore A23187 induced sperm acrosome reaction rate and sperm total acrosoin activity. Methods Sperm acrosome reaction was induced by calcium ionophore A23187,while sperm acrosin activity was determined by the modified Kennedy method. In this study, 150 cycles of IVF-ET were included, which were divided into Normal semen Group (85 cycles), Teratospermia Group (44 cycles), Asthenozoospermia Group (6 cycles). Compare the association between the calcium ionophore A23187 induced sperm acrosome reaction rate, total acrosin activity and normal morphology sperm percentage, concentration, activity in the 150 cycles and subgroups. Analyze the difference of the sperm acrosome reaction rate and total acrosin activity in Pregnancy Group and Non-pregnancy Group. Use the Receiver Operating Characteristic curve to make out the threshold of the sperm acrosome reaction rate and total acrosin activity for predicting the IVF pregnancy outcomes. Results In this 150 samples, there are no statistically significant differences between sperm acrosome reaction rate and sperm concentration, progressive velocity rate, the percentage of normal sperm morphology, fertility rate, and their related coefficient are respectively as follow:0.120, 0.018, 0.084, 0.054(P>0.05). But the negative correlation is statistically significant different between sperm acrosome reaction rate and teratozoospermia index(r=-0.183, P=0.025). There are significant correlations between sperm acrosin activity and sperm concentration(r=0.172, P<0.05), progressive velocity rate (r=0.306, P<0.05), sperm percentage normal morphology(r=0.222, P<0.05). There are no statistically significant differences between total sperm acrosin activity and teratozoospermia index(r=-0.027, P>0.05) and fertility rate(r=0.039, P>0.05). There are no statistically significant differences between sperm acrosome rate and fertility rate in Normal semen Group(r=-0.039, P>0.05), Teratospermia Group(r=0.185, P>0.05), Asthenozoospermia Group(r=-0.725, P>0.05). There are no statistically significant differences between total sperm acrosin activity and fertility rate in Normal semen Group (r=0.057, P>0.05), Teratospermia Group(r=-0.156, P>0.05), Asthenozoospermia Group(r=-0.319, P>0.05). The acrosome reaction rate is(23.65±16.60)%, which is significant higher than (17.96±11.00)%in Non-pregnancy Group(P=0.024). The total acrosin activity is (78.84±38.52)μIU/106 in Pregnancy Group, which is much higher than(64.72±31.67)μIU/106 in Non-pregnancy Group are statistically significant(P=0.031). The threshold value of acrosome reaction rate for predicting IVF pregnancy outcomes is 17.85%, and the sensitivity is 0.550, the specificity is 0.549. The threshold value of total acrosin activity for predicting IVF pregnancy outcomes is 59.84 IU/106, and the sensitivity is 0.662, the specificity is 0.570. Conclusion The value of calcium ionophore A23187 induce acrosome reaction rate to predict the male fertility and IVF outcomes are low. The acrosin activity may be a good predictor for male fertility, but not for IVF outcomes. The clinical value of calcium ionophore A23187 induced acrosome reaction may not be better than the total sperm acrosin activity.

Objective To explore the efficacy and safety of conjugated estrogens cream usage for cervical factor caused infertility. Method Forty two couples( one hundred ten cycles) were divided into two groups randomly. In control group,ovulation was documented by a transvaginal ultrasound scan prior to guide intercourse. In study group,put conjugated estrogens cream into vaginas of women. The cervical mucus score,thickness of endometrial and weather pregnancy were studied. Results There was significant differences in cervical mucus score and endometrial thickness between study group and control group. The total score of cervical mucus score was 6. 1636 ± 2. 0436in control group,but that of study group was 6364 ± 1. 6258; the difference was statistically significant( P < 0. 05).The thickness of endometrial was 6. 4 ± 1. 278mm in control group,but that of study group was 8. 4836 ± 0. 9527mm; the difference was statistically significant( P < 0. 05). There was no significant difference of pregnant rate in two groups( 11&19,P > 0. 05). Conclusion Conjugated estrogens cream can improve the cervical mucus score and endometrial thickness but could not increase pregnant rate distinctly.

Objective: To assess the relationship between the mixed agglutination reaction(MAR) test and outcome of in vitro fertilization-embryo transfer(IVF-ET) outcome.Methods: The clinical data of 562 infertile couples who underwent IVF treatment was analyzed by a retrospective study.According to the MAR test,the patients were divided into four groups: <10%,10%-30%,30%-50% and >50%.The fertilization rate,embryonic development and clinical pregnancy outcomes among the four groups were compared by statistical analysis.Results: A total of 22 cycles of total fertilization failure and 279 clinical pregnancies were obtained and the overall pregnancy rate was 51.7% per cycle.The maternal age,duration of infertility,total fertilization failure rate,sperm density,progressive motility,normal morphology,good-quality embryo rate,implantation rate,clinical pregnancy and abortion rate were not significantly different among these groups(P>0.05).However,the fertilization rate of the group with MAR>50% was significantly lower than other groups(P<0.05).Conclusion: The MAR test is valuable to predict the fertilization rate at IVF,however,the results of MAR test are not related to clinical pregnancy outcome.

目的 观察促性腺激素释放激素激动剂(GnRHa)促超排卵对子宫内膜转化生长因子-β(TGFβ1)蛋白表达的影响,及其与胞饮突和血清E2、P水平之间的相关性,探讨GnRHa方案低妊娠率的原因,为临床提高体外受精-胚胎移植(IVF-ET)妊娠率提供理论依据.方法 随机将96只雌鼠分成两组,自然周期组和控制性促超排卵组.应用扫描电镜观察子宫内膜胞饮突的表达,应用免疫组化SP法观察子宫内膜TGFβ1蛋白的表达,应用放射免疫法检测雌二醇(E2)、孕酮(P)水平.结果 GnRHa促超排卵可能通过改变血清E2、P水平,使子宫内膜TGFβ1蛋白表达高峰前移,使子宫内膜种植窗异常,导致子宫内膜发育与胚胎发育不同步,降低子宫内膜容受性,影响IVF-ET妊娠率.结论 GnRH促超排卵影响子宫内膜的植入窗.

OBJECTIVE:To investigate the effect of sperm acrosin activity on the IVF-ET outcome. METHODS:We analyzed sperm parameters, morphology and acrosin activity for 909 infertile husbands by computer-assisted self-assessment (CASA), modified Papanicolaou staining and N-alpha-benzoyl-DL-arginine-p-nitroanilide (BAPNA), respectively, and detected the rates of fertilization, cleavage, quality embryos, embryo cryopreservation, implantation, clinical pregnancy and abortion. The wives were identified as normal or with mere oviduct problems. RESULTS:The rate of normal sperm morphology and sperm motility, vitality, rapid progressive velocity and concentration were significantly lower in the abnormal acrosin activity group than in the normal one (P < 0.01). Significant positive correlations were observed between acrosin activity and the above-mentioned semen parameters (P < 0.01). There were no significant differences in the number of retrieved eggs, the rates of cleavage, quality embryos, embryo cryopreservation, non-embryo transfer cycles and miscarriages, and the number of transferred embryos between the two groups (P > 0.05). The fertilization rate, the percentage of transfer cycles with only 1 embryo and the rate of implantation and clinical pregnancy were notably higher in the normal acrosin activity group than in the abnormal one (P < 0.01). CONCLUSION:Sperm acrosin activity is closely related with semen parameters, and it helps to predict the sperm fertilizing capacity and IVF-ET outcome.

Objective: To investigate the relationship between sperm morphology and in vitro fertilization outcome. Methods: Sperm morphology of 480 infertile husband's semen was analyzed by modified Papanicolaou staining. The effect of sperm morphology onfertilization, cleavage, good quality embryo, embryo cryopreservation, implantation, pregnancy and abortion rates were evaluated. Results: Following with decrease of rate of normal morphology sperm, the concentration and motility of sperm were significantly decreased (P<0.01). IVF fertilization and good quality embryo rates in the group with high rate of normal morphology sperm(>15%) were significantly higher than those in the other two groups with low rate normal morphology sperm(≤4% or 5%-15%) (P<0.01). There were no differences in the cleavage, embryo cryopreservation, implantation, pregnancy and abortion rates among the three groups (P>0.05). Conclusion: Sperm morphology is involved in the IVF fertilization rate and embryo development potency. However, it could not be used topredict for IVF pregnant outcome.

Objective: To understand the sperm parameters before and after capacitation using Computer-Assisted Sperm Analysis (CASA). Methods: Using Paired-samples T test and Independent-samples T test, comparison of sperm CASA parameter before and after capacitation in 48 freeze-thawed and 116 fresh human sperm before AIH. Results: The sperm Linearity (LIN) in all groups, the sperm Straightness (STR) and elongation ratio in all except freezed group were reduced after capacitation (P<0.01); But the values of sperm Curvilinear velocity (VCL), Straight line velocity (VSL), average path velocity (VAP), amplitude of lateral head displacement (ALH) and Beat-cross frequency (BCF) were increased (P<0.01). The values of sperm VAP, LIN and elongation ratio in these who had fathered a child by AIH were higher than those who failed to father a child, but the sperm BCF and head area was significantly smaller on the contrary (P<0.05); The sperm motility (a+b) was no significance in two groups (P>0.05). Conclusion: The semen processing can improve the sperm parameters except LIN and STR in all groups. Successful AIH wasn't associated with sperm motility (a+b) in semen.

肾移植受者中人巨细胞病毒(HCMV)感染较为常见,其中10%～30%为术后有症状的HCMV感染[1].HCMV被膜磷蛋白65(HCMV-pp65)是HCMV DNA复制时即出现的一种抗原,在出现临床症状数日或1周前可检出HCMV-pp65阳性标本.我们采用间接免疫荧光染色法(IFA),检测肾移植术后病人外周血多形核细胞(PMNLs)中HCMV-pp65,以辅助临床对HCMV进行早期诊断与治疗.

Objective: To investigate the sensitivity of two bioassays for quality control of culture media used in in vitro fertilization (IVF) program. Method:The human sperm motility assay and the 2-cell mouse embryo bioassay were respectively performed to identify 20 batches commonly used in IVF program. Result: By using the sperm assay, 80% of media showed no sperm toxicity, while 50% of media had toxic effects on the development of 2-cell mouse embryo. There is a significant difference on detecting toxicity of medium between two bioassays(P0.05). Conclusion: The 2-cell mouse embryo bioassay would be more sensitive than the human sperm motility assay for quality control of culture media.

目的探讨精子膜表面AsAb导致男性不育的机制.方法对102例不孕夫妇的丈夫精液进行混合球蛋白反应试验(MAR),其中48例同时进行了血清AsAb的检测.结果48例同时进行MAR和ELISA的不育男性,精液MAR阳性率为14.6%(7/48);血清ELISA阳性率为50%(24/48).两种方法的阳性率有非常显著性差异(P＜0.001).102例进行MAR检测的不育男性中,随着精液量、密度、存活率、活动力、畸形率的变化,精子膜表面AsAb的阳性率均无显著性差异(P＞0.05).根据精子膜表面AsAb阳性与否分成两组组间的病人年龄、精液量、密度、存活率、活动力、畸形率均无显著性差异(P＞0.05).结论血清ELISA阳性率过高,不宜用于不育男性免疫因素的检测;精液精子膜表面AsAb并不影响精液分析的各项参数.因此,精液各项参数的分析对于男性免疫因素不育的筛查没有太大的意义.

目的探讨细胞因子基因多态性分布.方法采用PCR-SSP对58名随机人群的肿瘤坏死因子、转化生长因子、IL～6、IL-10、干扰素等细胞因子分别作基因多态性研究.结果肿瘤坏死因子三个基因型频率分别为0.431、0、431、0138,等位基因频率G=0.646、A=0.354;转化生长因子九个基因型频率分别是0.069、0.052、0.172、0.017、0.190、0.190、0.034、0.069、0.207,等位基因频率T=0.509、C=0491、C=0.276、G=0.724;IL-6三个基因型频率分别是0.204、0.638、0.155,等位基因频率C＝0.259、G=0.741;IL-10六个基因型频率分别是0.086、0.103、0.155、0.069、0.276、0.310,等位基因频率GCC=0.216、ACC=0.259、ATA=0.525;干扰素三个基因型频率分别是0.551、0.310、0.138,等位基因频率A=0.707、T=0.293.结论本组数据对细胞因子基因多态性研究有助于从分子水平去认识细胞因子在机体免疫调节的重要作用,对临床上某些疾病发病机理的研究、诊断和防治提供有力的依据.

目的应用原瓶传代培养的方法于经典的产前诊断的羊水培养方法中,以探讨建立提高培养成功率的方法.方法与结果于羊水培养10天左右作原瓶传代培养,实验组(原瓶传代)105例,培养成功率100%,对照组(经典培养方法)120例成功率92%.结论结果提示,原瓶传代培养可提高成功率至100%,并可获得满意的核型以供分析,确保了羊水产前诊断的准确性及临床应用性.

OBJECTIVE:To detect maternal serum fetal DNA for prenatal diagnosis of fetal sex.METHODS:Five to ten ml maternal serum was obtained from 24 pregnant women with 16 male fetuses and 8 female fetuses (including 4 carriers with X-linked diseases). The fetal sex was detected by analyzing the karyotype from amniotic fluid and villus. Serum DNA was extracted, nested polymerase chain reaction was employed the male fetal DNA.RESULTS:The sensitivity of the first polymerase chain reaction was 62.5%. The sensitivity of the first and the second reaction was 100%. There was no false positive. Fetal sex of 4 X-linked disease carriers was diagnosed. The determinate rate was 100%.CONCLUSION:Using fetal DNA from maternal serum and nested polymerase chain reaction was a quick, simple and accurate method for prenatal diagnosis of the fetal sex.

随着生殖助孕技术在人类医学中的蓬勃发展,发育遗传学及细胞工程在生殖学中日趋扮演重要的角色,多学科的互相渗透使生殖技术更加完善地为人类服务.二十一世纪第一期HUMAN REPRODUCTION以"Reproductive Genetics"代替了"Genetics and Genetic Diagnosis"作为一个新兴学科创建了遗传学中的新分支,开拓了"生殖遗传学"在生殖领域的研究及生殖医学中的新使命.