Infertility is a common health problem. For investigating the reason of infertility in the man, semen analysis plays very important role. Vitality stain is one of semen analysis. It is rare use for diagnosing and evaluating the result of infertility treatment. Our objectives: (1) To submit a fully worked-out of vitality staining technique. (2) To compare the effects of eosin and trypan blue staining for measuring live and death sperms. Methods: 52 semen samples were analyzed, compared the rate of death sperm base on the time after staining by eosin and trypan blue. Results: The average time for staining sperms by eosin: 16 seconds, by trypan blue: 19.67 seconds. The best process are: Mix 1 volume of eosin and 4-5 volume of semen, 1 volume of trypan blue and 2 volume of semen. Conclusions: (1) The more staining time, the more number of death sperm. (2) In vitality staining technique: The volume of semen should must be more than volume of stain chemical.
Spermatozoa, Human

In their embryology, Aristotle and Galen greatly disagreed on the role of human derived materials like menstrual blood and vaginal secretion (called by them female sperm or semen). This gap made those two ancients also disagree on their understanding of mother's role in the generation of the human body in her womb. During the Middle Ages, especially during the thirteenth century, the scholastics drew on those two ancient thoughts for some rational underpinnings of their philosophical and theological doctrines. However, the manners of adoption and assimilation were varied. For example, Albert the Great strived to reconcile the two in the image of Avicenna, one of the main and the most important sources of Galenist medicine in the thirteenth Century. By contrast, those scholastics who played an important role in the controversy over plurality/unicity of the substantial form, drew on their disagreements. For example, pluralists like Bonaventure, William of la Mare, and Duns Scotus appealed to Galenist medical perspective to underpin their positions and paved ways to decorate Virgin Mary's motherhood and her active contribution to the Virgin birth and to the manhood of her Holy Son. in contrast a unicist like Thomas Aquinas advanced his theory in line with Aristotelian model that Mary's role in her Son's birth and manhood was passive and material. Giles, another unicist, while repudiating Galenist embryology with the support of Averroes's medical work called Colliget, alluded to some theologically crucial impieties with which might be associated some pluralists' Mariology based on the Roman physician's model. In this processus historiae, we can see not only the intertwining of medieval medicine, philosophy, and theology, but some critical moments where medicine provided, side by side with philosophy, natural settings and explanations for religious marvels or miracles such as the Virgin birth, the motherhood of Mary, the manhood of Christ, etc. Likewise, we can observe two medieval maxims coincide and resonate: “philosophia ancilla theologiae” and “philosophia et medicina duae sorores sunt.”
Embryology ; Female ; Human Body ; Humans ; Parturition ; Philosophy ; Spermatozoa ; Theology

Many factors can result in male infertility, and a number of studies have indicated that reproductive difficulties are associated intimately with cytogenetic abnormalities. This article reviews studies on sperm chromosomes in infertile men and discusses the relationship between sperm chromosome abnormality and male infertility.
Chromosome Aberrations ; Chromosomes, Human ; Humans ; Infertility, Male ; genetics ; Karyotyping ; Klinefelter Syndrome ; genetics ; Male ; Spermatozoa ; ultrastructure

Repeated spontaneous abortion (RSA), with very complicated pathogenesis, has an incidence of about 0.5% 2.0%. As for the paternal part which provides half of the genes for the embryo, current studies mainly focus on the genes of somatic cells or germ cells. Chromosome abnormality of somatic cells, chromosome disorder of sperm, defects in sperm quality, genetic mutation, senility, infection and any other paternal gene abnormalities that affect the embryo would induce RSA. This paper presents an update on the above mentioned paternal factors which might result in RSA.
Abortion, Spontaneous ; etiology ; Chromosomes, Human, Y ; DNA Damage ; Female ; Humans ; Male ; Pregnancy ; Spermatozoa

Intrauterine insemination (IUI) together with controlled ovarian hyperstimulation (COH) has been increasingly used for the treatment of variety of subfertile indications, both male and female or even combined. The overall success rate of IUI ranges from 4% to 66%. The wide variance of success of the procedure is likely to be influenced by a number of factors. The pregnancy rate in the local setting has never been determined. This cross-sectional study reviewed all available clinical records of patients undergoing fertility work-up who had sperm processing in a hospital-based andrology unit and who underwent intrauterine insemination in either the hospital-based facility or a private clinic from January to December, 2004. Objective: It aimed to determine the pregnancy rate following IUI and assess the intrinsic and extrinsic variables affecting its success and describe the IUI's pregnancy outcome. The intrinsic factors include patient's age (male and female), number of subfertility years, previous reproductive history specifically involving the different factors (male, cervical, uterine, ovarian, tubal, peritoneal). Extrinsic factors include treatment effect and timing of IUI (medicine administered, monitoring of number and size of follicles, endometrial thickness, total motile count inseminated, number of inseminations) and preference for facility (hospital-based clinic or private clinics). Results: For the period of one year, there were a total of 1051 cycles of IUI, 305 in the hospital-based facility and 746 in private clinics. Due to limitation of accessible data, only 424 cycles were studied. However, out of the 424 cycles data retrieved, only 365 showed IUI outcomes. The overall pregnancy rate following IUI was 2.47%. In this study, it seems that only the wives' age (younger) and years of subfertility (2.9 years), were found to be associated with pregnancy rates. The median female age was 35.4 years (range 23.4-48.2), and median male age was 36.5 years (range: 25.0 - 54.4) with a median duration of subfertility of 6.0 years (range: 0.3 -18.0). Conclusion: There is no sufficient evidence to conclude that the other factors studied under treatment, different parameters and topography are associated with rates of pregnancy following IUI.

Human ; Male ; Female ; Adult ; Reproductive History ; Spouses ; Infertility ; Uterus ; Fallopian Tubes ; Insemination ; Fertility ; Spermatozoa

OBJECTIVETo study the technique of dual-color fluorescence in situ hybridization(D-FISH) and its application value in sperm chromosomes of Robertsonian translocation carriers.

METHODSThe technique of dual-color fluorescence in situ hybridization was used. Biotin labelled 13q14.3 specific probe and Digoxigenin labeled 14q11.1 specific probe were used for in situ hybridization of sperm specimens in 2 Robertsonian translocation carriers. Hybridization signals for chromosomes 13 and 14 in the sperm interphase nucleus were counted.

RESULTSUnder the microscope, Biotin labeled 13q 14.3 specific probe showed 1 green hybridization signal and Digoxigenin labeled 14q 11.1 specific probe showed 1 red hybridization signal. Interphase nucleus counter-stained with DAPI showed blue. From the total of 3,000 sperm interphase nuclei, the positive rate for 1 green hybridization signal and 1 red hybridization signal was 13q/14q(39.33%), for 1 green and 2 red was 13q/14q, 14q(11.57%), for 1 green was 13q/-(9.27%), for 2 green and 1 red was 13q,13q/14q(12.87%), for 1 red was -/14q(9.87%) and for 2 green and 2 red was 13q,13q/14q, 14q(12.26%).

CONCLUSIONSD-FISH of the human sperm interphase nucleus can be applied to the determination of sperm chromosomes of Robertsonian translocation carriers and the analysis of the laws of chromosomal segregation in the meiosis. The technique can be widely used in such aspects of human reproduction as insemination under the microscope and preimplantation embryos genetic diagnosis.

Adult ; Chromosomes, Human, Pair 13 ; Chromosomes, Human, Pair 14 ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Spermatozoa ; ultrastructure ; Translocation, Genetic

OBJECTIVETo analyze the numerical aberration of chromosome X, Y and 18 in the spermatozoa of asthenospermia patients by triple-color fluorescence in situ hybridization.

METHODSThe experiment included 10 asthenospermia patients and 5 healthy men with normal semen quality as controls. Fluorescence in situ hybridization (FISH) and probes for chromosomes including X, Y and 18 were used to determine the frequency of the aneuploid of the chromosomes in spermatozoa.

RESULTSOf the 45,547 spermatozoa counted from the semen samples, the hybridization rate was 99.18%. The frequencies of the chromosome disomies including XX18, XY18, YY18, X1818 and Y1818 were (0.124 +/- -0.086)%, (0.360 +/- 0.380)%, (0.109 +/- 0.195)%, (0.342 +/- 0.746)% and (0.299 +/- 0.564)% in the case group and (0.014 +/- 0.019)%, (0.090 +/- 0.080)%, (0.030 +/- 0.031)%, (0.068 +/- 0.103)% and (0.075 +/- 0.083)% in the control. The sperm aneuploid rate was 9.25% in the former and 2.70% in the latter, with significant difference in between (P< 0.01).

CONCLUSIONAsthenospermia patients have a higher aneuploid rate of sperm chromosome than normal fertile men. However, larger samples are yet to be studied to obtain more scientific evidence.

Aneuploidy ; Asthenozoospermia ; genetics ; Chromosome Painting ; methods ; Chromosomes, Human, Pair 18 ; Chromosomes, Human, X ; Chromosomes, Human, Y ; Humans ; Male ; Sex Chromosome Aberrations ; Spermatozoa ; metabolism

The acrosome reaction is a Ca(2+)-dependent exocytotic process that is a prerequisite step for fertilization. External calcium entry through voltage-activated Ca(2+)channels is known to be essential in inducing the acrosome reaction of mammalian spermatozoa. Due to their complex geometry, however, electrophysiological identification of sperm Ca(2+)channels has been limited. Here we identified Ca(2+)channel mRNAs expressed in motile human sperm using RT-PCR and their levels were compared using RNase protection assays. L-type, non- L-type, and T-type Ca(2+)channel mRNAs were detected by RT-PCR using degenerate primers. Cloning and sequencing of the PCR products revealed alpha1B, alpha1C, alpha1E, alpha1G, and alpha1H sequences. RT-PCR using specific primers repeatedly detected alpha1B, alpha1C, alpha1E, alpha1G, and alpha1H mRNAs, and additionally alpha1I mRNA. But alpha1A and alpha1D messages were not detected. Relative expression levels of the detected Ca(2+)channel subtypes were compared by RNase protection assays. The abundance of detected mRNA messages was in the following order: alpha1H> or =alpha1G> or =alpha1E> or =alpha1B>alpha1C>alpha1I. These findings indicated that human motile sperm express multiple voltage-activated Ca(2+)channel RNAs among which T-type and non-L-type channel messages are likely to be predominantly expressed. Based on their relative expression levels, we propose that not only T-type but also non-L-type calcium channels may be major gates for the external calcium influx, required for the acrosome reaction.
Calcium/*metabolism ; Calcium Channels/biosynthesis/classification/*genetics ; Human ; Male ; RNA, Messenger/*metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Spermatozoa/*metabolism

AIMTo simultaneously determine the localization of histones and protamines within human sperm nuclei.

METHODSImmunofluorescence of the core histones and protamines and fluorescence in situ hybridization of the telomere region of chromosome 16 was assessed in decondensed human sperm nuclei.

RESULTSImmunofluorescent localization of histones, protamine 1 (PRM1) and protamine 2 (PRM2) along with fluorescence in situ hybridization localization of chromosome 16 telomeric sequences revealed a discrete distribution in sperm nuclei. Histones localized to the posterior ring region (i.e. the sperm nuclear annulus), whereas PRM1 and PRM2 appeared to be dispersed throughout the entire nucleus.

CONCLUSIONThe co-localization of the human core sperm histones with the telomeric regions of chromosome 16 is consistent with the reorganization of specific non-protamine regions into a less compacted state.

Cell Nucleus ; metabolism ; Chromosomes, Human, Pair 16 ; metabolism ; Histones ; metabolism ; Humans ; Male ; Protamines ; metabolism ; Spermatozoa ; metabolism ; Telomere ; metabolism

AIMTo determine if Yq microdeletion frequency and loci of deletion are similar in two tissues (blood and sperm) of different embryological origin.

METHODSThe present study included 52 infertile oligozoospermic cases. In each case, DNA was isolated from blood and sperms and polymerase chain reaction (PCR) microdeletion analysis was done from genomic DNA isolated from both the tissues. The PCR products were analyzed on a 1.8% agarose gel. PCR amplifications found to be negative were repeated at least three times to confirm the deletion of a given marker.

RESULTSOnly 1 case harbored microdeletion in blood DNA, whereas 4 cases harbored microdeletion in sperm DNA.

CONCLUSIONThe frequency of Yq microdeletions is higher in germ cells as compared to blood. As the majority of infertile couples opt for assisted reproduction procreation techniques (ART), Yq microdeletion screening from germ cells is important to understand the genetic basis of infertility, to provide comprehensive counseling and most adapted therapeutics to the infertile couple.

Chromosomes, Human, Y ; genetics ; DNA ; blood ; genetics ; isolation & purification ; Humans ; Male ; Repetitive Sequences, Nucleic Acid ; Sequence Deletion ; Spermatozoa ; physiology