Analysis of 12 kinds of cytokines in seminal plasma by flow cytometry and their correlations with routine semen parameters.

Objective: To investigate the levels of 12 kinds of cytokines in seminal plasma and their correlations with routine semen parameters.

Methods: The remaining seminal plasma samples of 134 patients undergoing routine semen examination were collected for detecting cytokines. The parameters for sperm concentration, percentage of progressively motile sperm (PR), and motility were analyzed by a computer-assisted sperm analysis (CASA) system.

Hyperlipidemia is negatively associated with pregnancy outcomes in patients following frozen-thawed embryo transfer: A retrospective study.

Background And Aims: It is demonstrated that lipid metabolism disorders are associated with the reproductive performances of assisted reproductive technology. However, it is little known whether hyperlipidemia is associated with the endometrial receptivity and pregnancy outcomes of patients undergoing frozen-thawed embryo transfer (FET).

Methods: This was a retrospective analysis involving 554 infertile women undergoing FET.

Effect of hyperlipidemia on the outcome of fertilization in non-obese patients with polycystic ovary syndrome.

Introduction: It is little known whether hyperlipidemia alone has adverse effects on the outcome of fertilization (IVF) in patients with polycystic ovarian syndrome (PCOS).

Methods: The PCOS patients with body mass index (BMI) < 30 kg/m were performed IVF or intracytoplasmic sperm injection treatment, including 208 fresh cycles and 127 frozen embryo transfer (FET) cycles. All the patients were divided into hyperlipidemia and control groups, and embryo quality and pregnancy outcomes between the two groups were compared.

Performance evaluation of sperm concentration, motility, and morphological analysis for GSA-810 series of sperm quality analysis system.

Background: The performance evaluation of each computer-assisted sperm analysis (CASA) system may provide a basis for the interpretation of clinical results and further improvement of the CASA system.

Methods: The accuracy of the GSA-810 CASA system was evaluated by detecting latex bead quality control products. The precision of sperm concentration, morphology, and percentages of progressively motile sperm (PR) were evaluated by coefficient of variation (CV).

Analysis of selected sperm samples by a computer-assisted system with high frame rate: A prospective study.

Background And Aims: Due to the rapid motility of the selected sperm, sperm parameters cannot be accurately determined by the manual method. So, the application of a computer-assisted sperm analysis system with a high frame rate (HFR-CASA, 85 Hz) in sperm selection is investigated.

Methods: A total of 177 semen samples were collected for sperm selection with density gradient centrifugation.

Correlations of joint detection of 22 vaginal microbes with routine examination results of vaginal secretions and assisted reproductive outcomes.

The correlations of joint detection of 22 vaginal microbes with routine examination results of vaginal secretions and assisted reproductive outcomes were investigated. There were 37 samples with abnormal vaginal microecology in 107 vaginal secretion samples. The top 5 detection rates of microorganisms were Ureaplasma urealyticum (73.

Investigation on the mechanisms of human sperm DNA damage based on the proteomics analysis by SWATH-MS.

Background: Spermatozoa have the task of delivering an intact paternal genome to the oocyte and supporting successful embryo development. The detection of sperm DNA fragmentation (SDF) has been emerging as a complementary test to conventional semen analysis for male infertility evaluation, but the mechanism leading to SDF and its impact on assisted reproduction remain unclear. Therefore, the study identified and analyzed the differentially expressed proteins of sperm with high and low SDF.

Effects of six kinds of sperm staining methods on human sperm size and evaluation of their staining effects.

Background: Large- and small-headed sperm are common morphological abnormalities. If different sperm staining methods affect sperm size, they will make a difference in the accuracy of sperm morphological analysis results. In this case, the normal reference values of sperm head parameters for different staining methods should be established.

Screening of sperm antigen epitopes by phage display technique and its preliminary clinical application.

Background: At present, there is a lack of standardized preparation methods of sperm antigen for the detection of antisperm antibody (AsAb). To screen sperm antigen mimotopes from a phage display random peptide library and use them to establish an enzyme-linked immunosorbent assay (ELISA) for the detection of AsAb, immunoglobulins were extracted from the sera of rabbits with positive AsAb and negative AsAb, respectively, by the saturated ammonium sulfate method, and a phage display 12-mer peptide library was affinity panned by the extracted immunoglobins coated on the ELISA plate. Then, the obtained positive phage clones were identified by ELISA and sent for sequencing and peptides synthesis.

Investigation of the mechanisms leading to human sperm DNA damage based on transcriptome analysis by RNA-seq techniques.

Research Question: What are the molecular mechanisms leading to human sperm DNA damage?

Design: Semen samples were collected and the sperm DNA fragmentation index (DFI) was assessed. Differentially expressed RNA in spermatozoa with a high (DFI ≥30%, experimental group) or normal (DFI <30%, control group) DFI were identified by RNA-sequencing (RNA-seq) technology, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was performed. Three differentially expressed RNA related to sperm DNA damage and repair, namely PMS1, TP53BP1 and TLK2, were validated using real-time quantitative (RT-qPCR).

High DNA stainability (HDS) should not be recommended as a marker for the detection of sperm DNA damage.

There was a marker of high DNA stainability (HDS) in the detection of sperm DNA damage, which was defined as the sperm with high green stainability (HIGRN). The sperm with normal double-stranded DNA was stained green by acridine orange (AO). However, the sperm with high green fluorescence or HDS were thought of as immature sperm or the sperm with poor chromatin condensation.

[Standardization and quality control for detection of sperm DNA damage by flow cytometry: A preliminary investigation].

Objective: To investigate preliminarily the standardization and quality control for the detection of sperm DNA damage by flow cytometry.

Methods: Semen samples were randomly selected and observed for the effects of acid denaturation time, acridine orange (AO) staining time, semen sample refrigeration, freezing and repeated freezing-thawing on the results of sperm DNA fragmentation index (DFI).

Results: Sperm DFI increased gradually with the prolongation of acid denaturation time, significantly at 2 minutes in comparison with that at 30 seconds (P<0.

[Establishment and evaluation of a flow cytometry technique reflecting the severity of human sperm DNA damage].

Objective: To establish a flow cytometry (FC) technique reflecting the severity of human sperm DNA and evaluate its performance.

Methods: We analyzed the red and green fluorescence peaks of normal sperm in 1 165 human semen samples, defined the average lower limit of 5% of green fluorescence and the average upper limit of 95% of red fluorescence as the red and green fluorescence limits of the four-quadrant gate, and established an FC technique for detection of the sperm DNA fragmentation index (DFI), analysis of its repeatability and linear range and determination of the reference value of normal fertile men. We also analyzed the correlations of the sperm DFI, mild DNA damage marker (DFIm) and severe DNA damage marker (DFIs) with sperm concentration and motility in 122 men from the Infertility Clinic of Zhongda Hospital.

[Automatic detection and clinical application of semen biochemical markers].

Human seminal plasma is rich in potential biological markers for male infertility and male reproductive system diseases, which have an application value in the diagnosis and treatment of male infertility. The methods for the detection of semen biochemical markers have been developed from the manual, semi-automatic to the present automatic means. The automatic detection of semen biochemical markers is known for its advantages of simple reagent composition and small amount of reagents for each test, simple setting of parameters, whole automatic procedure with few errors, short detection time contributive to batch detection and reduction of manpower cost, simple calibration and quality control procedure to ensure accurate and reliable results, output of results in the order of the samples in favor of clinical diagnosis and treatment, and open reagents applicable to various automatic biochemistry analyzers.

Identification and preliminary study of immunogens involved in autoimmune prostatitis in human males.

Background: Experimental models have confirmed that autoimmunity is an important factor in the onset of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS); however, there is no conclusive evidence on whether autoimmune prostatitis exists in human males.

Methods: Rabbits were immunized with either human prostate tissue homogenates or normal saline and the antiserum was collected. Two-dimensional electrophoresis (2-DE) was performed on the homogenates and Western blotting was conducted on the sera.

[Association of abnormal length of Y chromosome with semen quality and outcome of assisted reproductive technology in humans].

Objective: To investigate the association of the abnormal length of human Y chromosome with semen quality and the outcome of assisted reproductive technology (ART).

Methods: Based on the karyotype, we assigned the patients undergoing ART to a normal control, a long Y chromosome (Y>18), and a short Y chromosome group (Y<22). We compared the semen parameters and numbers of embryos and high-quality embryos among the three groups of patients and performed statistical analysis of the obtained data using Chi-square distribution and t-test.

Analysis of human sperm DNA fragmentation index (DFI) related factors: a report of 1010 subfertile men in China.

Background: Many factors may lead to sperm DNA damage. However, it is little known that the correlations of sperm DNA damage with obesity-associated markers, and reproductive hormones and lipids levels in serum and seminal plasma.

Methods: In our prospective study, a total of 1 010 subfertile men, aged from 18 to 50 years old, were enrolled from August 2012 through June 2015.

Relationship Between Amyloid Precursor Protein in Seminal Plasma and Abnormal Penile Sympathetic Skin Response in Lifelong Premature Ejaculation.

Introduction: Hyperactivity of the sympathetic nervous system can play an important role in lifelong premature ejaculation (PE). Our previous study found that amyloid precursor protein (APP) levels in seminal plasma of patients with PE were clearly increased. Amyloid-β (Aβ) is derived from APP.

[Animal models of autoimmune prostatitis and their evaluation criteria].

Chronic prostatitis is a highly prevalent disease of unclear etiology. Researches show that autoimmune reaction is one cause of the problem. An effective animal model may help a lot to understand the pathogenesis and find proper diagnostic and therapeutic strategies of the disease.

Accuracy Evaluation of The Depth of Six Kinds of Sperm Counting Chambers for both Manual and Computer-Aided Semen Analyses.

Background: Although the depth of the counting chamber is an important factor influencing sperm counting, no research has yet been reported on the measurement and comparison of the depth of the chamber. We measured the exact depths of six kinds of sperm counting chambers and evaluated their accuracy.

Materials And Methods: In this prospective study, the depths of six kinds of sperm counting chambers for both manual and computer-aided semen analyses, including Makler (n=24), Macro (n=32), Geoffrey (n=34), GoldCyto (n=20), Leja (n=20) and Cell-VU (n=20), were measured with the Filmetrics F20 Spectral Reflectance Thin-Film Measurement System, then the mean depth, the range and the coefficient of variation (CV) of each chamber, and the mean depth, relative deviation and acceptability of each kind of chamber were calculated by the closeness to the nominal value.

[Correlations of 24 biochemical markers in seminal plasma with routine semen parameters].

Objective: To investigate the correlations of 24 biochemical markers in the seminal plasma with routine semen parameters.

Methods: According to the WHO5 standards, we analyzed the routine semen parameters of 66 subfertile men, including the semen volume, sperm concentration, total sperm count, sperm motility, and the percentage of progressively motile sperm (PR). Based on the calibration and quality control measures and using an automatic biochemistry analyzer or electrolyte analyzer, we detected 24 biochemical markers in the seminal plasma of the patients, including total protein (TP), albumin (Alb), globulin (Glb), uric acid (UA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP), γ-glutamyltransferase (GGT), lactate dehydrogenase (LDH), creatine kinase (CK), alpha hydroxybutyrate dehydrogenase (αHBDH), adenosine deaminase (ADA), glucose (Glu), triglyeride (TG), total cholesterol (TC), urea nitrogen (UN), creatinine (Cr), high-sensitive C-reactive protein (hsCRP), K+, Na+, Cl- , Ca, Mg, and phosphorus (P).

Relationship between Lipids Levels of Serum and Seminal Plasma and Semen Parameters in 631 Chinese Subfertile Men.

Objective: This prospective study was designed to investigate the relationship between lipids levels in both serum and seminal plasma and semen parameters.

Methods: 631 subfertile men were enrolled. Their obesity-associated markers were measured, and semen parameters were analyzed.

A Pilot Comparative Study of 26 Biochemical Markers in Seminal Plasma and Serum in Infertile Men.

Introduction: The relationships of the biochemical components in seminal plasma and serum, and their origins and physiological effects in male reproductive system have been poorly understood.

Methods: Based on the calibration and quality control measures, 26 biochemical markers, in seminal plasma and serum samples from 36 male infertility patients with nonazoospermia were detected and compared.

Results: Only PA was undetectable in all seminal plasma samples.

[Related factors of sperm DNA damage: Advances in studies].

The detection of sperm DNA damage, as an important supplement to semen routine examination strategies, has been applied in some clinical andrology laboratories. What factors may lead to sperm DNA damage remains one of the concerns among many andrologists. Present studies show a variety of factors of sperm DNA damage, including age, environmental pollutants such as organophosphorus and organochloride pesticides, plasticizer, heavy metals such as lead, carcinogens such as polycyclic aromatic hydrocarbons (c-PAHs) and zearalenone (ZEA), male reproductive system diseases or systemic diseases such as varicocele, infection, tumor, spermatogenesis and maturation dysfunction, spinal cord injury and endocrine disorders, seasons and temperature, lifestyle, abstinence time, semen refrigeration, semen handling in vitro, and certain medications.

Annexin V-induced rat Leydig cell proliferation involves Ect2 via RhoA/ROCK signaling pathway.

This study investigated the effect of annexin V on the proliferation of primary rat Leydig cells and the potential mechanism. Our results showed that annexin V promoted rat Leydig cell proliferation and cell cycle progression in a dose- and time-dependent manner. Increased level of annexin V also enhanced Ect2 protein expression.