Evaluating an external quality assurance program for semen analysis in China during 2009-2020.

INTRODUCTION: Semen analysis (SA) plays a key role in guiding treatments of male reproductive diseases and infertility due to male factors; however, it remains challenging to conduct an accurate SA due to lack of standardization, highly subjective assessments, and problems with automated procedures. Therefore, quality assurance (QA) and teaching courses are essential for making the laboratory results more consistent. MATERIALS AND METHODS: The external quality assurance (EQA) scheme was organized by national human sperm bank technology training bases in Guangdong province in China between 2009 and 2020. Until 2020, 124 laboratories from China participated in the EQA program. The EQA scheme per year has been organized involving two semen aliquots for sperm concentration, two video recordings for motility, and two smears for sperm morphology. All samples used in the EQA scheme were obtained from different healthy donors or patients. RESULTS: We estimated that the median coefficient of variation (CV) of sperm concentration, ignoring the method used, was 26.6%. Using a 100 µm deep counting chamber led to a decreasing CV of 13.6%. For sperm motility, the median CV of nonprogressive motility was high (50.8%), but the CV of progressive motility (13.2%), immotile sperm (14.3%), and total motility (11.8%) were acceptable. The morphology assessment revealed large variability (44.4%) irrespective of the classification criteria. DISCUSSION: The reduction of interlaboratory variability is still a challenge during SA in China. Therefore, it is critical to increase awareness of joining EQA schemes and establish standardized training centers to follow WHO-recommended procedures toward Chinese standards.

[Causes of failure in autologous sperm preservation in the human sperm bank and countermeasures].

OBJECTIVE: To analyze the causes of failure in autologous sperm cryopreservation (ASCP) in the human sperm bank and propose some countermeasures to improve the success rate of ASCP and promote it in human sperm banks. METHODS: We retrospectively analyzed the reasons for and causes of failure in ASCP 1 156 males undergoing ASCP in the Human Sperm Bank of Guangdong Province from January 1, 2016 to December 31, 2019. RESULTS: Of the 1 156 cases of ASCP, 857 (74.1%) succeeded and 299 (25.9%) failed, with a failure rate of 56.1% (96/171) in the microdissection testicular sperm extraction (micro-TESE) group, 29.9% (67/224) in the reproduction insurance group, 21.2% (27/100) in the non-tumor disease group and 17.2% (109/525) in the tumor group, with statistically significant difference between the four groups (χ2 = 109.926, P < 0.01). The causes of failed ASCP included failure to extract semen (14.0% ï¼»42/299ï¼½), failure to meet the standard of sperm cryopreservation (67.6% ï¼»202/299ï¼½), giving up ASCP for low semen quality (7.4% ï¼»22/299ï¼½), and giving up ASCP for some other reasons (11.0% ï¼»33/299ï¼½), including worry about the reliability of cryopreservation technology (6.0% ï¼»18/299ï¼½), suspicion about the complexity of the ASCP procedures (3.0% ï¼»9/299ï¼½) and expectation for fertility recovery after chemotherapy withdrawal (2.0% ï¼»6/299ï¼½). CONCLUSIONS: In view of different causes of failure in ASCP, human sperm banks can provide individualized cryopreservation schemes, including guidance with masturbation or the use of sperm extraction instrument in sperm extraction, strengthening sperm preservation-related education and the awareness of reproduction protection and earlier sperm preservation among cancer patients, promoting the cryopreservation of microsamples of motile sperm by microsample or single sperm freezing, and development of testis tissue cryopreservation to preserve the fertility of children with cancer.

[Differentially expressed long non-coding RNAs in spermatozoa with different fertilizing abilities].

OBJECTIVE: To search for conception capacity-related long non-coding RNAs (lncRNA) and explore their possible roles in fertilization. METHODS: We obtained 10 semen samples, 5 of high and the other 5 of low fertilizing ability, extracted large RNAs, established a cDNA library, and performed RNA sequencing with the HiSeq 2000 sequencing system. Using the bioinformatics method, we assembled and predicted lncRNAs, screened differentially expressed genes between the two groups by NOIseq, analyzed the lncRNAs with the box plot and volcano plot, and determined their expression patterns by hierarchical cluster analysis. We examined the functional classification of differentially expressed lncRNAs by pathway and gene ontology (GO) enrichment and predicted those of some lncRNAs by lncRNA-mRNA interaction analysis and intersection analysis with up- and down-stream cis-acting elements. RESULTS: A total of 147 1615 lncRNAs were identified in all the semen samples, including 463 596 novel ones and 8 019 known ones, with 4 052 differentially expressed lncRNAs, 985 upregulated and the other 3 067 downregulated. Box plot and volcano plot filtering analyses showed statistically significant differences in the expressions of the lncRNAs between the two groups, and so did hierarchical cluster analysis. GO functional annotations manifested the involvement of the differentially expressed lncRNAs in the metabolic process, biological regulation, membrane and organelle formation, and protein-nucleotide binding. Pathway analysis showed that the differentially expressed lncRNAs were related to transport and catabolism, cell motility, signaling molecular interactions, signaling transduction, and signaling pathways in the development and immune systems. The functions of the 5 lncRNAs predicted were shown to be associated with sperm motility, acrosomal reaction and signal transduction during fertilization. CONCLUSIONS: Differentially expressed lncRNAs may play an important role in fertilization and become biomarkers for the assessment of sperm quality.

[Antiatherogenic effect of piperlonguminine on experimental atherosclerosis in rabbits].

OBJECTIVE: To investigate of antiatherogenic effect and possible mechanisms of piper longuminine. METHOD: The atherosclerotic model was established by the hypercholesterol feeding rabbits. Male Mew Zealand rabbits were randomly divided into five groups: normal group, model group, the high-dose (5 mg x kg(-1) x d(-1)) and low-dose (2.5 mg x kg(-1) x d(-1) group of piperlonguminine, and simvastatin group (5 mg x kg(-1) x d(-1)). All the rabbits were fed for 60 days. Blood samples were taken from the ear edge vein of rabbits in the day before the experiment, and in the days of 20, 40 and 60 days after the experiment, respectively. All the rabbits were fasted for at least twelve hours before the blood was taken. The blood serum were analyzed for total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C). The blood serum of the 60th day were also analyzed for superoxide dismutase (SOD), malondialdehyde (MDA) and nitric oxide (NO). At last, the pathological observation of aorta and heart samples were carried out. RESULT: Compared with those in model group, the TC, TG and LDL-C levels were reduced (P < 0.05) and the HDL-C was raised in the piperlonguminine group; also, the serum SOD and NO level was raised (P < 0.05), MDA level was reduced in the piperlonguminine group (P < 0.05). Area percentage of aorta plaque was reduced (P < 0.01) in the piperlonguminine group. The aorta and heart injury was abated and coronary artery angusty extent was markedly abatement (P < 0.01). The results of observation through transmission electron microscope (TEM) indicated that the fine structure of aortal pathological degree was markedly abated. CONCLUSION: The piperlonguminine could inhibit the atherogenesis formation and development, which might be due to regulating the lipid metabolism and enhancing the antioxidation.

[Correlation between biological characteristics of volunteer donors and sperm parameters].

OBJECTIVE: To explore the correlation between the biological characteristics of volunteer donors and sperm parameters. METHODS: Sperm parameters were analyzed for 778 primary volunteer donors by CASA system with standard methods recommended by WHO including sperm concentration, motility rate, VCL, VSL, ALH, LIN, STR and BCF, and their biological characteristics were recorded. RESULTS: Of all the volunteer donors, students accounted for more than a third, 89.2 percent were under the age of 35 years, 71.6 percent between 166 and 175 cm in height, 91.7 percent of normal weight and 77.1 percent with college education. The median sperm concentration and motility rate were 107.00 x 10(6)/ml and 75 percent respectively. Age was weakly correlated with such semen indexes as concentration (r = 0.210, P = 0.000) , motility rate (r = 0.213, P = 0.000), volume (r = 0.165, P = 0.002), VAP (r = 0.259, P = 0.000), VSL (r = 0.281, P = 0.000), VCL (r = 0.190, P = 0.000), BCF (r = 0.243, P = 0.000) and LIN (r = 0.192, P = 0.000). The semen indexes of the 26-35 age group were a little better than those of the 20-25. Height showed no correlation to semen indexes. Education and occupation were somewhat related with other sperm parameters (P < 0.01) than ALH (P = 0.695/0.886). CONCLUSION: Age, occupation and education bear some correlation with sperm parameters except ALH, while height has none with any of them.