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1.
人卵泡液诱导的顶体反应、精子形态与体外受精率的关系   总被引:4,自引:2,他引:4  
目的:探讨人卵泡液诱导的顶体反应、精子形态及体外受精率3者之间的相互关系。方法:应用Spearm an法对79对不育患者的人卵泡液诱导的顶体反应、精子形态(采用结晶紫染色及精子形态严格分类标准)及体外受精率等指标进行等级相关分析。结果:人卵泡液诱导的顶体反应率与正常形态精子百分率之间存在极显著正相关(n=49,r=0.376 3,P<0.01),但人卵泡液诱导的顶体反应率与体外受精率以及正常形态精子百分率与体外受精率之间均未见显著相关性(n分别为21和50,r分别为0.266 6和0.001 8,P均>0.05)。结论:人卵泡液诱导的顶体反应率与正常形态精子百分率之间存在极显著正相关,但两者与体外受精率之间均未见显著相关性。  相似文献   

2.
The aim of this study was to evaluate the effect of date seed oil (DSO) on epididymal sperm characteristics and testicular antioxidant enzymes in male mice. DSO was diluted into isotonic saline solution (0.9%) and different doses (5, 10, 15 and 20%) were prepared. Fifty male mice were divided into five groups; in four groups DSO was given by intraperitoneal injection of oil solution for 28 days. The control group was injected by isotonic saline solution without DSO. Body and reproductive organ weights, sperm characteristics (count, motility, viability and morphology) were assessed. In addition, levels of malondialdehyde (MDA), activities of superoxide dismutase (SOD) and catalase (CAT) were investigated in testes. A significant increase in sperm count, motility and viability of all treated animal groups was observed when compared with the control group ( P  < 0.05). Unlike, the percentage of abnormal sperm was significantly lower in all treated groups than in the control group ( P  < 0.05). A significant decrease in MDA levels and marked increase in SOD and CAT activities in mice treated with high doses of DSO (15 and 20%) were also noted. We suggest that DSO can improve the epididymal sperm quality and could ameliorate the testicular strategy defences.  相似文献   

3.
The study consisted of application of anti-ubiquitin antibodies (Abs)-coated iron oxide-nanoparticles (IONPs) for minimisation of oxidative stress to contemporary live spermatozoa from the raw semen. Round-shaped IONPs (12.09 ± 0.91 nm) after two-stage functionalisation (silanisation and pegylation) were conjugated with Abs. Four aliquots from each of the 24 ejaculates (4 buffalo bulls) formed Control (Group I) and treatment (II, III and IV) groups; each containing 150 ± 25 million dead/damaged spermatozoa. IONPs-Abs complex were added at ratio of 1:1 (0.5 µg/ml), 1:2 (1.0 µg/ml) and 1:4 (2.0 µg/ml), respectively, in Groups II, III and IV. The semen quality parameters showed improvement at lag-stage (post-nano-purification before processing for cryopreservation). The mean post-thaw motility (%) in Group IV was found to be greater (p < .05) than Group I. Moreover, the overall DNA integrity (%) at post-thaw stage was improved in the nano-purified semen samples. The value of malondialdehyde was greater (p < .001) in Group I than Groups II, III and IV. The mean total antioxidant capacity and superoxide dismutase (U/mg protein) activity values in Group IV was greater (p < .05) than Group I. The study results show that IONPs conjugated with anti-ubiquitin Abs at 2.0 µg/ml can be an effective dose for depletion of dead/damaged spermatozoa from buffalo ejaculates to minimise oxidative stress.  相似文献   

4.
刺五加水提物体外对弱精子症患者精子运动参数的影响   总被引:4,自引:1,他引:3  
目的:研究不同浓度的刺五加水提物体外对弱精子症患者精子运动的影响,探讨其可能的作用机制。方法:将35例弱精子症患者经手淫法获得并通过上游优化处理的精子,与不同浓度刺五加水提物共同孵育30、60、120、180 m in后,应用计算机辅助精子分析系统(CASA)观察不同浓度(2.5、5、10、20 g/L)刺五加水提物对人精子各运动参数的作用。结果:不同浓度刺五加水提物能明显提高弱精子症患者精子的运动能力,在浓度为5 g/L和10 g/L时,精子活率(MOT)、前向运动精子百分率、曲线运动速度(VCL)、直线运动速度(VSL)和平均路径速度(VAP)与空白对照组相比,差异均有显著性(P<0.05)。结论:刺五加水提物在体外能明显改善弱精子症患者精子运动能力,其最佳浓度为10 g/L。  相似文献   

5.
Semen samples of 24 patients were analysed. Volumes were measured and the numbers of progressively motile (PMS), motile (MS) and nonmotile spermatozoa (NMS) were determined. These 24 samples appeared to show a large variation in motility percentages and numbers. Spermatozoa of these semen samples were isolated from the seminal plasma and exposed to induced radical oxygen stress imposed by iron/ascorbate. Lipid peroxidation (LPO) was quantified as thiobarbituric acid reactive material. The contributions of PMS, MS and NMS were also estimated. It was found that the PMS did not contribute to the formation of lipid peroxides. The cellular radical defence system of PMS may offer them adequate protection against the harsh conditions of radical oxygen stress. Stepwise regression analyses showed that only the population of NMS contributed significantly to the explanation of the variance in LPO production (R2 = 0.56, P < 0.001). Pre-existing membrane lipid peroxides were not detected in spermatozoa. It is therefore suggested that LPO takes place only after radical oxygen stress has exhausted the cellular defence system. LPO is not the initial, but one of the later, events leading to the death of spermatozoa. It is concluded that the population of progressively motile spermatozoa in semen samples does not contribute to the production of thiobarbituric acid reactive substances as induced by in vitro radical oxygen stress.  相似文献   

6.
In order to select sperm characteristics that can predict the outcome of in-vitro fertilization-embryo transfer (IVF-ET), semen samples delivered in conjunction with this treatment were studied carefully. We have analysed these data retrospectively in relation to the outcome of treatment. Ninety-one couples were treated for tubal infertility by IVF-ET. Fifteen women became pregnant. Sperm were isolated from semen using a swim-up technique and in most cases 40-80 x 10(3) (range 20-120 x 10(3)) motile sperm per ovum were used for insemination. The couples were divided into three groups: group A who achieved pregnancies, group B who achieved cleaved ova but no pregnancies, and group C who achieved no ova that were cleaved 48 h after oocyte recovery. Comparisons between these groups showed that some characteristics of the native semen samples and the swim-up preparations were significantly different: the sperm concentration (P = 0.001) and total sperm count (P = 0.003) in the native sample, the number of sperm recovered during 30 min of swim-up (P = 0.001), and the specific progressive motility of sperm in the swim-up preparation, both at the time of insemination and on each day, up to 5 days thereafter (P = 0.002-0.028). No pregnancy was achieved with a sperm concentration below 26 x 10(6) ml-1 in the native sample. Some of the sperm characteristics studied in this paper may be of value in the pretreatment evaluation for IVF treatment.  相似文献   

7.
Summary. In order to correlate functional and biochemical parameters, frozen bovine sperm samples presenting diverse motility were studied. Specimens collected from four bulls (A, B, C and D) were graded according to motile spermatozoa percentage and vigour, regarding the sample from bull A as highest motility, those from bulls B and C of intermediate motility and that from bull D of low motility. In order to evaluate lipid peroxidation enhanced by the addition of sodium ascorb-ate and ferrous sulphate, thiobarbituric acid reactive substances (TEARS) were measured. Values were 0.34 ± 0.18 and 4.95 ± 0.31 nmol TEARS/108 spermatozoa for samples A and D, respectively. Superoxide dismutase (SOD) activity was 144±16.48 and 44±4.0 IU/1010 spermatozoa for A and D, respectively, while intermediate values were recorded for samples B and C. The capability to undergo acrosome reaction induced by calcium ionophore A23187 was significantly lower for sample D, but differences were negligible for the remaining three samples inter se. Motile spermatozoa percentage correlated closely both with SOD activity ( r = 0.92) and with TEARS production ( r =−0.80), but not with acrosome reacted percentage ( r =0.65). From correlation data, it would be inferred that motility is not entirely indicative of sperm quality.  相似文献   

8.
《Renal failure》2013,35(5):612-617
Hexachlorobutadiene (HCBD) is a potent nephrotoxin in rodents. Pharmacological studies have shown that pomegranate fruit preparations have antioxidant, anti-inflammatory chemopreventive effects. In this study, the effect of pomegranate seed oil (PSO) on HCBD-induced nephrotoxicity was investigated in adult male rats. Animals were divided into five groups. Group 1 was treated with corn oil (1 mL/kg, i.p.). Group 2 received a single dose of HCBD (50 mg/kg, i.p.). Groups 3–5 were treated with PSO (0.16, 0.32, and 0.64 mg/kg, i.p., respectively) 1 h before HCBD (50 mg/kg, i.p.) injection. A significant elevation of serum creatinine and urea (p < 0.001) levels as well as urine glucose and protein (p < 0.001) concentrations (as markers of acute renal failure) was observed 24 h after administration of HCBD as compared to control group. HCBD also caused a significant decrease in total thiol content (p < 0.001) and a significant increase in thiobarbituric acid reactive species (TBARS, as an index of lipid peroxidation) levels (p < 0.001) in kidney homogenate samples. PSO pretreatment resulted in a significant and dose-dependent decrease in serum creatinine (p < 0.001) and urea levels (p < 0.001) as well as urine glucose (p < 0.001) and protein concentrations (p < 0.001) when compared with HCBD treated alone. PSO also significantly reversed the HCBD-induced depletion in total thiol content (p < 0.001) and elevation in TBARS (p < 0.001) in kidney homogenate samples. The results of this study showed that PSO clearly attenuated HCBD-induced nephrotoxicity, but explanation and mechanism of this protection need further explorations.  相似文献   

9.
Progesterone (P4) is crucial for the physiological function of spermatozoa. In the study, we investigated the correlation between P4‐induced sperm acrosome reaction (AR) and parameters including sperm progressive motility, normal morphology and sperm DNA fragmentation (SDF), and compared the in vitro fertilization (IVF) predictive values of these indicators based on the multivariate regressions analysis and receiver operator characteristics (ROC) curve analyses. The results demonstrated a negative correlation between P4‐induced sperm AR and the SDF, with the correlation ?9.05 (?17.25 to ?0.84), p<0.05, n = 47). No relationship was found between the sperm progressive motility, normal morphology and the induced AR. The P4‐induced AR and SDF were both significantly correlated to the fertilization rate. ROC curve analyses indicated that P4‐induced AR was a better prognostic predictor for the fertilization rate compared with the SDF, with the areas under the curve 0.729 (0.580–0.849), p<0.01 and 0.637 (0.484–0.772), p=0.16 respectively. The cut‐off value for P4‐induced AR to predict “50% fertilization rate” was 23.4% with sensitivity and specificity of 63.3% and 88.2% respectively. The overall results indicated that the assessment of P4‐induced AR seemed to be a more sensitive indicator for fertilization rate in vitro compared with other sperm parameters.  相似文献   

10.
目的:研究壬基酚和镉离子在体外对小鼠精子顶体反应(AR)的影响。方法:从小鼠的输精管获得精子,体外培养使精子获能,加30μmol/L的A23187诱导精子顶体反应,然后使用不同浓度的壬基酚(10、20、30、60、100μmol/L),或者镉离子(500、2500、5000μmol/L)处理,对照组使用相应的载体溶剂处理。用FITC-PSA荧光染色法分析精子顶体反应。结果:当壬基酚浓度<30μmol/L时,小鼠精子顶体反应率与对照组比较没有显著差异(P>0.05),而当壬基酚浓度>60μmol/L时能够显著地抑制小鼠精子顶体反应发生率(P<0.01),并且观察到精子存活率随着壬基酚浓度增加而降低。与壬基酚作用不同,用镉离子对小鼠精子进行处理,在所选浓度内(500~5 000μmol/L)均对精子顶体反应无显著影响(P>0.05),且精子存活率与镉离子浓度变化无关。结论:壬基酚与镉离子对小鼠精子发生的作用是通过不同的途径来实现的,前者可以直接抑制顶体反应,而后者则与精子顶体反应无关。  相似文献   

11.
Cinnamon and its contents have multifactorial properties such as antioxidant, anti‐inflammatory and antidiabetic. Male infertility is one of the major health problems in life. The aim of this study was to investigate the effects of long‐term cinnamon bark oil (CBO) ingestion on testicular antioxidant values, apoptotic germ cell and sperm quality of adult rats. Twelve male healthy Wistar rats were divided into two groups, each group containing six rats. While olive oil was given to control group, 100 mg kg?1 CBO was administered to the other group by gavage daily for 10 weeks. Body and reproductive organ weights, sperm characteristics, testicular lipid peroxidation and antioxidant enzyme activities, and testicular apoptosis via terminal deoxynucleotidyl transferase–mediated dUTP nick end labelling (TUNEL) method were examined. A significant decrease in malondialdehyde level and marked increases in reduced glutathione level, glutathione peroxidase and catalase activities were observed in rats treated with CBO compared with the control group. CBO consumption provided a significant increase in weights of testes and epididymides, epididymal sperm concentration, sperm motility and diameter of seminiferous tubules when compared with the control group. However, CBO consumption tended to decrease the abnormal sperm rate and apoptotic germ cell count, but it did not reach statistical significance. It is concluded that CBO has improvement effect on testicular oxidant–antioxidant balance and sperm quality, and its consumption may be useful for asthenozoospermic men.  相似文献   

12.
目的:优化电刺激取精术获取的精子处理和冷冻复苏方法,为临床不射精症的治疗和生育力保存提供新途径。方法:将20例同房不射精患者电刺激获取的精液(或尿液)样本分为两组,其中6例采用PBS洗涤并用Modified-HTF重悬(A组),另外14例用Modified-HTF直接洗涤并重悬(B组),对处理后精子行CASA,分析精子浓度和活动率;采用麦管载体和一步熏蒸法冷活动率为(1.12±0.41)%,M-HTF处理组的精子活动率为(9.23±9.43)%;冷冻复苏后,A组精子复苏后活动率和冷冻复苏率分别为(0.24±0.41)%、8.37%;B组的精子活动率和复苏率显著增加,分别为(4.11±2.33)%、35.41%。同样,电刺激尿液样本中,PBS处理组的精子活动率为(1.30±0.51)%,M-HTF处理组的精子活动率提高至(5.48±5.32)%;冷冻复苏后,A组的精子活动率和复苏率分别为(0.26±0.40)%、6.31%;B组的精子活动率和复苏率,分别为(3.02±1.64)%、45.88%。结论:本研究所建立的电刺激精子经体外培养及冷冻复苏的方法,可以显著增加精液和尿液样本中精子的活力,改善精子的冷冻复苏率,为临床不射精症的治疗和生育力保存提供新途径。  相似文献   

13.
The aim of this study was to compare the effectiveness of antioxidants including cysteamine (2.5, 7.5 mm ), hyaluronan (0.25, 1 mg ml?1) and fetuin (5, 10 mg ml?1) in the freezing of Brown Swiss bull semen. The best percentages of CASA motilities were achieved with 10 mg ml?1 of fetuin and 2.5 mm of cysteamine. For sperm morphology, 10 mg ml?1 of fetuin and 2.5 mm of cysteamine had better protective effects (P < 0.001). The results of hypo‐osmotic swelling test showed that the percentage values of membrane integrity in all the groups, excluding that supplemented with 5 mg ml?1 of fetuin, were higher than those of the control group (P < 0.001). Results obtained for the DNA damage of sperm cells demonstrated that 0.25 mg ml?1 of hyaluronan, and 2.5 and 7.5 mm of cysteamine led to lower rates of spermatozoa with damaged DNA, compared with the control group (P < 0.001). The maintenance of superoxide dismutase and glutathione peroxidase antioxidant activities following freeze‐thawing with 2.5 and 7.5 mm of cysteamine and 10 mg ml?1 of fetuin was demonstrated to be at a higher level in comparison with the control group (P < 0.001). Malondialdehyde formation was found to be lower in the groups supplemented with 0.25 mg ml?1 of hyaluronan and 7.5 mm of cysteamine after the freeze‐thawing process (P < 0.001).  相似文献   

14.
In this study, it was aimed to investigate the likelihood of detrimental effects of carbon tetrachloride (CCl4) on male reproductive system through oxidative stress mechanism and also protective effects of cinnamon bark oil (CBO). For this purpose, 28 healthy male Wistar rats were divided into four groups, seven rats in each. Group 1 received only olive oil daily; group 2 was treated with 100 mg kg?1 CBO daily; group 3 was treated with only 0.25 ml kg?1 CCl4 weekly; and group 4 received weekly CCl+ daily CBO. All administrations were made by intragastric catheter and maintained for 10 weeks. Body and reproductive organ weights, sperm characteristics, testicular oxidative stress markers and testicular apoptosis were examined. CCl4 administration caused significant decreases in body and reproductive organ weights, testicular catalase (CAT) activity, sperm motility and concentration, and significant increases in lipid peroxidation (LPO) level, abnormal sperm rate and apoptotic index along with some histopathological damages compared with the control group. However, significant improvements were observed in absolute weights of testis and epididymis, all sperm quality parameters, LPO level, apoptotic index and testicular histopathological structure following the administration of CCl4 together with CBO when compared to group given CCl4 only. The findings of this study clearly suggest that CBO has protective effect against damages in male reproductive organs and cells induced by CCl4.  相似文献   

15.
目的:评估短时受精联合早期补救性卵细胞胞质内单精子注射(ICSI)术在完全受精失败周期的应用结局。方法:10例体外受精-胚胎移植(IVF-ET)周期获取的卵细胞随机分组,分别采用短时受精与过夜受精,比较受精率与胚胎质量。105例常规IVF-ET周期短时受精后,行极体观察,早期判断完全受精失败后行补救性ICSI术,报道临床结局。结果:3h短时受精与过夜受精在受精率与胚胎质量上无明显差异(P>0.05)。短时受精联合早期受精失败补救后,临床妊娠率为53.3%,种植率为38.0%,出生婴儿64名。结论:短时受精联合早期补救性ICSI对常规IVF-ET周期完全受精失败患者的临床结局有明显改善。  相似文献   

16.
Background: The present study was conducted to elucidate the protective role of Withania somnifera against bromobenzene induced nephrotoxicity and mitochondrial dysfunction in rats. Methods: In this study, Wistar albino rats of either sex were divided into six groups consisting of six animals each. The first one was control, those in group II received bromobenzene (10?mmol/kg, intragastric intubation) once, but group III and IV animals received W. somnifera (250 and 500?mg/kg, orally), respectively for 8 days followed by bromobenzene once on the 8th day and silymarin (100?mg/kg, orally) was given for 8 days to group V animals and then bromobenzene on the last day. Group VI animals received only W. somnifera (500?mg/kg) for 8 days. Results: The levels of renal lipid peroxidation, lysosomal enzymes and glycoprotein were increased significantly (p?<?0.05) in the bromobenzene alone treated rats and antioxidant status and mitochondrial enzymes were found to be decreased, when compared to the control group. The levels of kidney functional markers (urea, uric acid and creatinine) were also found to be abnormal in serum of bromobenzene alone treated rats. On the other hand, administration of W. somnifera (250 and 500?mg/kg) along with bromobenzene offered a significant dose-dependent protection to the biochemical alterations as observed in the bromobenzene alone treated rats, which was also evidenced by histopathology. Conclusion: Thus, the oral administration of W. somnifera significantly protected against the bromobenzene induced nephrotoxicity and renal mitochondrial dysfunction in rats.  相似文献   

17.
Dr.  H. H. PUSCH 《Andrologia》1987,19(5):514-527
Motility behaviour of spermatozoa in 289 ejaculates was investigated by laser-Doppler-spectroscopy in the andrology-laboratory of the University Hospital for Gynecology and Obstetrics, Graz/Austria. Conventional spermiograms, performed on the same ejaculates, permitted comparisons between estimated motility and laser-measurements, comprising the following parameters: Total sperm motility, mean sperm velocity, percentages of progressive motile spermatozoa and mean progressive velocity. In order to elucidate the importance of sperm motility for the fertilization of oocytes in vivo and in vitro, ejaculates of six different groups of patients were evaluated. In vivo: 173 patients from the andrologic outpatient service (group A); 38 patients from the andrologic outpatient service, whose wives became pregnant (group B) and 20 donors with recently proved fertility (group C). In vitro: 8 patients, whose ejaculates failed to fertilize the oocytes (group D); 39 patients with successful fertilization (group E) and 12 patients with ongoing, clinical pregnancy of their wives after ivF (group F). In addition, motility behaviour was assessed in native ejaculates 24 hours after ejaculation, as well as changes in sperm motility after capacitation. Statistical evaluations of group comparisons revealed a good correlation between estimated results and laser measurements. Mean sperm velocity and progressive sperm motility are the most selective parameters within the group differences in oligozoospermic ejaculates. Besides motility, morphology is of importance in vivo as well as in vitro. Based on the results of measurements obtained in the different groups, the following guidelines for standard values in laser-Doppler-spectroscopy are proposed: Total sperm motility greater than 53%; mean sperm velocity greater than 42 microns/sec.; progressive sperm motility greater than 34% and mean progressive velocity greater than 68 micron/sec. If these values are not reached, medical treatment is recommended.  相似文献   

18.
目的 :研究首乌还精胶囊含药血清体外对人精子运动能力和受精能力的影响。 方法 :通过人精子与首乌还精胶囊含药血清共培养 ,观察首乌还精胶囊对人精子运动能力 (精子运动CASA分析 )、精子顶体反应和穿透去透明带仓鼠卵能力的影响。 结果 :含药血清加入共培养可显著提高人精子运动速度 [精子运动路径速度(VAP)、精子轨迹速度 (VCL)、精子前向运动速度 (VSL) ](P <0 .0 1)、精子头部侧置振幅 (ALH)和鞭毛摆动频率(BCF)及前向运动精子密度 (P <0 .0 5 ) ,精子顶体反应发生率 (P <0 .0 0 1) ,卵子受精率 (FR)及受精指数 (FI) (P <0 .0 1)。且含药血清刺激作用呈现出一定的量效关系。 结论 :首乌还精胶囊具有提高人精子运动能力和受精能力的作用。  相似文献   

19.
Purpose: Oxidative stress due to hyperglycemia is a major cause of diabetes complications. The aim of this study was to evaluate the effects of pomegranate seed oil (PSO) on serum biochemical parameters, cardiomyopathy and nephropathy induced by diabetes mellitus.

Method: W/A adult rats were divided into four groups (12 each): group 1, received saline (1?mL/kg), group 2, received streptozotocin (STZ, 65?mg/kg, a single dose as i.p.), groups 3 and 4, received STZ?+?PSO (0.4 and 0.8?mL/kg, daily by gavage, respectively). After three weeks, six rats of each group and one week later the remaining animals were anesthetized, blood samples were taken for measuring serum biochemical parameters. Sections of heart and kidneys were used for histopathological studies and the remaining tissues were homogenized for measuring malondialdehyde (MDA) and total sulfhydryl groups.

Results: Significant elevation of serum creatinine and urea, LDL, triglyceride, glucose levels as well as urine markers, MDA levels in tissue homogenates and a significant decrease in total thiol content and serum HDL were observed in STZ-treated group as compared with control group. PSO treatment resulted in a significant decrease in tissue MDA content, serum creatinine and urea levels as well as urine markers as compared with STZ-treated group. Lipid profile was ameliorated with PSO treatment. PSO also significantly reversed STZ-induced depletion in thiol content and histological abnormality. Effect of PSO was more specific at 28th than 21th days of study.

Conclusion: The results showed that PSO has a protective effect against diabetes complications in rats.  相似文献   

20.
Summary.  The proportion of spermatozoa that undergo spontaneous acrosome reaction in vitro is relatively low. The proportion can be enhanced by incubation with either biological inducers such as follicular fluid or chemicals like calcium ionophore. It has been suggested that improper acro-somal reaction may be a cause of fertilization failure in vitro. The objectives of the present study were to assess the acrosomal status of human sperm following follicular fluid or calcium ionophore treatment and to analyse the relationship between spontaneous and induced acrosome reaction and fertilization rates in vitro by standard in vitro fertilization (IVF) technology. In all, 53 semen samples (22 normal and 31 subnormal) were studied. The effect of calcium ionophore A 23187 and follicular fluid was assessed using the fluorescence activated cell sorter. IVF results were evaluated in relation to the acrosome status of the sperm samples. Our results demonstrate that the effect of follicular fluid on the acrosomal status correlated positively with the effect obtained by the calcium ionophore (Pearson's correlation r = 0.45). A significantly higher percentage of maximal acrosome change ( P <0.02) was found in cases where fertilization occurred (19/27), than in sperm samples that did not achieve fertilization in vitro (8/27). The present finding that follicular fluid induced acrosome reaction can serve as a predictive tool which is as good as the ionophore treatment for assessing IVF outcome, supports the use of this method for clinical purposes.  相似文献   

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