Type 2 diabetes mellitus and the risk of male infertility: a Mendelian randomization study.

Objective: To assess the causal effect of type 2 diabetes mellitus (T2DM) on male infertility (MI) and erectile dysfunction (ED) by Mendelian randomization (MR) analysis. Methods: Data for T2DM, MI, and ED were obtained from genome-wide association studies (GWAS) involving 298, 957, 73, 479, and 223, 805 Europeans, respectively. We performed univariate MR analysis using MR Egger, Weighted median (WM) and Inverse variance weighted (IVW) methods to assess causal effects among the three. Through the Genotype Tissue Expression (GTEx) database, single-nucleotide polymorphisms (SNPs) that affect the expression levels of T2DM-related genes were located using expression quantitative trait loci (eQTL). Results: MR analysis showed a significant causal relationship between T2DM and ED (WM, OR: 1.180, 95%CI: 1.010-1.378, P = 0.037; IVW, OR: 1.190, 95%CI: 1.084-1.300, P < 0.001). There is also a significant causal relationship between T2DM and MI (MR Egger, OR: 0.549, 95%CI: 0.317-0.952, P = 0.037; WM, OR: 0.593, 95%CI: 0.400, P = 0.010; IVW, OR: 0.767, 95%CI: 0.600-0.980, P = 0.034). ED may not cause MI (P > 0.05). We also found that rs6585827 corresponding to the PLEKHA1 gene associated with T2DM is an eQTL variant affecting the expression of this gene. Conclusion: T2DM has a direct causal effect on ED and MI. The level of PLEKHA1 expression suppressed by rs6585827 is potentially associated with a lower risk of T2DM.

Ectopic pregnancy and failed oocyte retrieval during in vitro fertilization stimulation: Two case reports.

BACKGROUND: Due to a slight rise in beta-human chorionic (ß-hCG) levels that are undetectable, and vaginal bleeding that is similar to regular menstruation, ectopic pregnancy (EP) that occurs during the expected menstrual cycle prior to ovulation induction as part of in vitro fertilization (IVF) is likely to be undiagnosed. We present two cases of unexpected EP and emphasize the importance of the ß-hCG assay when an unexplained increase in progesterone is present prior to the triggering of ovulation during controlled ovarian stimulation (COS). CASE SUMMARY: A 26-year-old woman with primary infertility and a 31-year-old woman with secondary infertility. Both patients sought IVF treatment due to fallopian tube obstruction and underwent COS using the gonadotropin-releasing-hormone (GnRH)-antagonist protocol. In the late stage of COS, progesterone levels in both patients significantly increased, and luteinizing hormone levels decreased, followed by oocyte retrieval failure. A right salpingectomy was performed and tubal ectopic pregnancy was diagnosed by pathology in the first patient, and the second patients was diagnosed with a suspected EP abortion because her ß-hCG levels declined to 12.5 mIU/mL. After full recovery for 2 mo, the first patient entered a new IVF treatment cycle with a GnRH-antagonist regimen and successfully achieved eight oocytes and three viable embryos. After 6 mo, the second patient received another COS treatment with a progestin-primed ovarian stimulation protocol and successfully achieved nine oocytes and five viable embryos. CONCLUSION: ß-hCG levels in the initial and midterm phases of COS must be considered in patients with unusual hormone dynamics.

Association between transferred embryos and multiple pregnancy/live birth rate in frozen embryo transfer cycles: A retrospective study.

Background: Physicians need an appropriate embryo transfer strategy to address the challenge of reducing multiple birth rates, while maintaining the couples' live birth rate during assisted reproductive technology. Methods: We included 10,060 frozen embryo transfer cycles from January 2015 to March 2020 in reproductive medical center of Ruijin hospital, Shanghai, China. Patients were grouped according to the number and grade of cleavage-stage embryo or blastocysts transferred. Live birth rate and multiple live birth rate were compared among groups of women of different ages. Multivariable logistic regression models were used to estimate the risk of multiple live birth using different combinations of transferred embryos. Results: The transfer of double good-quality embryos was an independent predictor for multiple birth in women aged <30 years and those aged 36-39 years [<30 years: aOR =1.54 (95% CI: 1.14-2.06, P < 0.01); 36-39 years: aOR =1.84 (95% CI: 1.0-3.4, P < 0.01)]. Further, for women aged <36 years, the transfer of good-quality + poor-quality blastocysts was an independent predictor for multiple birth rate [<30 years: aOR=2.46 (95% CI: 1.45-4.18, P < 0.01); 31-35 years: aOR =4.45 (95% CI: 1.97-10.06, P < 0.01)]. Conclusions: Single-good-quality blastocyst transfer is recommended for women of all ages. When good-quality cleavage embryos are available, the choice of single or double embryo transfer with good- or average-quality embryo should depend on the age of women. Double embryo transfer with the highest possible grade of embryos is recommended for women aged ≥40 years.

Sperm DNA fragmentation in Chinese couples with unexplained recurrent pregnancy loss.

We aimed to study the association between sperm DNA fragmentation and recurrent pregnancy loss (RPL) in the Chinese population via a retrospective observational study of Chinese couples who had experienced RPL between May 2013 and August 2018. The study population included 461 men from couples with RPL and 411 men from a control group (couples with clinical pregnancy via in vitro fertilization owing to female causes). Routine semen analysis, sperm chromatin analysis, and microscopic (high-power) morphological analysis were performed using semen samples. Semen samples were assessed for volume, sperm count, and motility. The sperm DNA fragmentation index (DFI) was calculated, and the median DFI was obtained. Men were categorized as having normal (37.8%; DFI ≤ 15.0%), moderate (33.6%; 15.0% < DFI < 30.0%), or severe (28.6%; DFI ≥ 30.0%) DNA fragmentation levels. The percentage of men with severe DNA fragmentation was significantly higher in the RPL (42.3%) group than that in the control group (13.1%), whereas the percentage of men with normal levels of DNA fragmentation was significantly lower in the RPL group (22.8%) than that in the control group (54.7%). Subsequent analysis also demonstrated that the sperm DNA fragmentation rate had a moderate reverse correlation with the sperm progressive motility rate (r = -0.47, P < 0.001) and the total motile sperm count (r = -0.31, P < 0.001). We found a positive correlation between RPL and sperm DNA fragmentation. The results suggest that increased sperm DNA damage is associated with RPL.

Tumor-infiltrating regulatory T cells are positively correlated with angiogenic status in renal cell carcinoma.

BACKGROUND: Immune cells within a tumor microenvironment have shown modulatory effects on tumor angiogenic activity. Renal cell carcinoma (RCC) is a hypervascular tumor that reportedly increases the frequency of regulatory T cells (Tregs) in tumor tissues. This study investigated the correlation between Tregs infiltration and angiogenic status in RCC. METHODS: Thirty-six patients with RCC were enrolled in the present study, and twenty age-matched healthy donors were included as the control. Tregs were defined as CD4(+)CD25(high)CD127(low/-) T cells. The frequency of Tregs in peripheral blood and tumor infiltrating lymphocytes (TILs) were determined by flow cytometry. The expression of vascular endothelial growth factor (VEGF) in surgical resection specimens were measured with a commercial enzyme-linked immunosorbent assay (ELISA) kit. Microvessel density (MVD) was calculated on slides stained with CD34 antibody. Spearman's rank correlation was performed to evaluate the correlation between the frequencies of Tregs in TILs and VEGF values, as well as between frequencies of Tregs and MVD determinations. RESULTS: Compared to healthy controls, the frequency of peripheral blood Tregs was significantly increased in patients with RCC (P < 0.05). The percentage of tumor-infiltrating Tregs was higher than that of peripheral blood Tregs in patients with RCC (P < 0.01). In addition, the frequency of tumor-infiltrating Tregs was shown to significantly correlate with the pathological stage (P < 0.05) and nuclear grade (P < 0.01). Importantly, a significant positive correlation was observed between the frequency of tumor-infiltrating Tregs and VEGF protein expression (r = 0.51, P < 0.05), as well as between frequencies of Tregs and MVD score (r = 0.39, P < 0.05). CONCLUSIONS: These observations suggest that the high pro-angiogenic status of RCC may be associated with the accumulation of Tregs in the local microenvironment. Angiogenesis networks may be connected with immune tolerance units and cooperate with each other to facilitate tumor growth and progression.

Sperm chromatin structure assay results after swim-up are related only to embryo quality but not to fertilization and pregnancy rates following IVF.

The aim of this study was to investigate whether the sperm chromatin structure assay (SCSA) results after swim-up are related to fertilization rates, embryo quality and pregnancy rates following in vitro fertilization (IVF). A total of 223 couples undergoing IVF in our hospital from October 2008 to September 2009 were included in this study. Data on the IVF process and sperm chromatin structure assay results were collected. Fertilization rate, embryo quality and IVF success rates of different DNA fragmentation index (DFI) subgroups and high DNA stainability (HDS) subgroups were compared. There were no significant differences in fertilization rate, clinical pregnancy or delivery rates between the DFI and HDS subgroups. However, the group with abnormal DFI had a lower good embryo rate. So, we concluded that the SCSA variables, either DFI or HDS after swim-up preparation, were not valuable in predicting fertilization failure or pregnancy rate, but an abnormal DFI meant a lower good embryo rate following IVF.

Distribution of serum prostate-specific antigen in Chinese healthy men: a population-based study.

BACKGROUND: The morbidity and mortality of prostate cancer have been increasing rapidly in recent China. There were few studies investigating prostate-specific antigen (PSA) values ranges in the healthy Chinese population. We performed this study to determine the distribution of serum PSA in a large healthy Chinese population. METHODS: From January 2001 to May 2008, 11 150 healthy Chinese men aged 30 - 79 years came to our hospital for routine health check-up. All subjects without a previous diagnosis of prostate cancer, a history of prostate surgery, or urogenital tract infection were proposed to undergo systematic serum PSA measurement and digital rectal examination (DRE). Men with normal DRE and PSA ≤ 4.0 ng/ml and those PSA > 4.0 ng/ml or abnormal DRE but without adverse findings on prostate biopsy were included (n = 9358). Age and serum PSA concentration were recorded and correlated through Logistic regression analysis. RESULTS: The 95th percentile serum PSA concentration was 1.89 ng/ml for men aged 30 to 39 years, 2.19 ng/ml for men aged 40 to 49 years, 2.88 ng/ml for men aged 50 to 59 years, 4.42 ng/ml for men aged 60 to 69 years, and 6.52 ng/ml for men aged 70 to 79 years. The serum PSA concentration correlated with age (P < 0.0001) with an annual increase of 0.97% for men in 40 years, 1.58% for men in 50 years, 3.04% for men in 60 years, and 3.99% for men in 70 years. CONCLUSIONS: The serum PSA level correlates directly with age in Chinese men older than 40 years, not in Chinese men younger than 40 years old. Chinese men have lower PSA level compared with white men above 60 years of age, not in those under 60 years of age.

A sperm viability test using SYBR-14/propidium iodide flow cytometry as a tool for rapid screening of primary ciliary dyskinesia patients and for choosing sperm sources for intracytoplasmic sperm injection.

Spermatozoa viability tests based on dual-color flow cytometry after staining with Sybr-14/propidium iodide were performed on 44 men with complete asthenospermia for primary ciliary dyskinesia (PCD) screening, and seven were identified with PCD by electron microscopy of ultrastructural ciliary defects. Six PCD patients underwent eight intracytoplasmic sperm injection therapy cycles using ejaculated sperm or testicular sperm, obtaining a mean fertilization rate of 46.6%, with three healthy babies born and one in utero at the time of writing.

[Impact of cigarette smoking on human sperm DNA integrity].

OBJECTIVE: To investigate the influence of cigarette smoking on human sperm DNA integrity. METHODS: Totally, 784 cases of male infertility were selected from our case database and grouped according to whether they were smokers or nonsmokers, how much they smoked (< or = 10, 11-19 and > or = 20 cigarettes/d) and how long they smoked (< or = 5, 6-9 and > or = 10 yr). Sperm DNA integrity was measured using sperm chromatin structure assay (SCSA) and flow cytometry. DNA fragmentation and immature spermatozoa were expressed by the DNA fragmentation index (DFI) and high DNA stainability (HDS) respectively. Conventional sperm parameters and sperm DNA integrity were compared among different groups. RESULTS: The total semen volume and percentage of grade a + b sperm were lower and the sperm morphological abnormality was higher in the > or = 20 cigarettes/d and > or = 10 yr groups than in the others (P < 0.05). DFI and HDS were significantly higher in the smokers than in the nonsmokers (P < 0.05). HDS was negatively correlated with the percentage of grade a + b sperm (r = -0.18, P < 0.05) and both DFI and HDS were positively correlated with the rate of sperm malformation (r = 0.31 and r = 0.39, P < 0.05). CONCLUSION: Smoking more than 20 cigarettes a day or longer than 10 years has deleterious effects on the semen volume, percentage of grade a + b sperm and sperm morphology of the smokers. Cigarette smoking decreases sperm DNA integrity and nuclear maturation.

[A 10-year survey on birth defects after in vitro fertilization-embryo transfer in Shanghai].

OBJECTIVE: To survey birth defects of neonates conceived by using various types of in vitro fertilization and embryo transfer (IVF-ET) between 1998 and 2007 in Shanghai. METHODS: From 1998 to 2007, 8507 neonates from 6551 pregnancies conceived through assistant reproductive technology (ART) from 7 reproductive medicine center in Shanghai were enrolled in this retrospective study, including Shanghai Ji-Ai Genetics and IVF Institute, Shanghai Jiaotong University School of Medicine affiliated Renji Hospital, Ruijin Hospital, China Welfare Institute International Maternal and Infant Health Hospital, Shanghai First Maternity and Infant Health Hospital, Shanghai the Ninth People's Hospital and the Second Military Medical University affiliated Changhai Hospital. The clinical data about the type and incidence of birth defect were analyzed. Meanwhile, the factors associated with birth defect were investigated, such as various ART, gender, mother age, numbers of gestational sac, the source and quality of sperm. RESULTS: The birth defect rate was 1.08% (92/8507) and the most remarkable malformation occurred in circulation system [34% (31/92)]. The rates of major congenital malformations were 1.21% (34/2799) in fresh conventional IVF-ET, 1.07% (20/1871) in IVF-frozen-thawed embryo transfer (IVF-FET), 1.04% (23/2212) in fresh intracyto plasmic sperm injection (ICSI)-ET and 0.92% (15/1625) in ICSI-FET, which did not show statistical difference (P > 0.05). There was also no significantly different incidence of malformations between 1.12% (49/4371) in male neonates and 1.02% (42/4136) in female neonates (P > 0.05). However, the occurrence of congenital malformation is positively related with maternal age, the rates were 0.84%(41/4884) in mother's age less than 30 years and 1.77% (16/902) in mother' age more than 35 years, which reached statistical difference (P < 0.05). It also showed remarkable different incidence among 0.53% (25/4679) in singletons, 1.59% (57/3576) in twins and 3.97% (10/252) in triplets (P < 0.05). No remarkable difference of malformation rate among sperm sources used in fertilization were observed, including 1.09% (81/7419) in ejaculated sperm, 1.21% (7/579) in percutaneous epididymal aspiration (PESA) and 0.79% (4/509) in donor sperm (P > 0.05). CONCLUSIONS: The overall incidence of birth defect in neonates from ART is similar to those conceived naturally and is not associated with different types of IVF-ET, embryo cryopreservation, sperm sources. However, maternal age and multiple pregnancies confer the higher possibility of birth defect.

[Factors related to occurrence of twin pregnancy after double-embryo transfer in vitro fertilization cycles].

OBJECTIVE: To analysis high risk factors of twin pregnancy after double-embryo transfer in fresh in vitro fertilization-embryo transfer (IVF-ET) cycles. METHODS: From Jan. 2003 to Dec. 2007, 275 infertile cases underwent IVF-ET or intracytoplasmatic sperm injection (ICSI) and obtained clinical pregnancy in Reproductive Medical Center, Ruijin Hospital affiliated to Shanghai Jiaotong University. A total of 280 cycles were performed, which were classified into single pregnancy group (198 cycles) and twin pregnancy group (82 cycles). The general information, patient and embryo characteristics were compared between those two groups, then univariate and multivariate regression were analyzed. RESULTS: (1) There was no statistical difference in the following clinical features between single and twin pregnancy groups, such as patients ages, the ratio of secondary infertility, period and possible causes of infertility (P > 0.05). (2) When comparing basal level of follicle stimulating hormone (FSH), mean numbers of follicles, mean obtained ovum, ovarian responsibility (ratio of follicle stimulation hormone dose/number of oocyte retrieved), endometrial thickness given by human chorionic gonadotropin (hCG), no significant difference were observed between two groups (P > 0.05). Twin pregnant group had fewer cycles of in vitro fertilization treatment when compared with single pregnancy group (0.18 +/- 0.16 vs. 0.22 +/- 0.21, P = 0.03). (3) No significant difference was observed in the following clinical index, including fertilization approaches, mean numbers of embryo, mean score of transferred embryo, developmental stage of top quality embryo, morphological score of embryo, morphological score of the second best embryo transferred (P > 0.05). The number of top-quality embryos and the development stage score of the second best embryo transferred were higher than those of single pregnant group (3.8 +/- 3.3 vs. 2.9 +/- 2.5, P < 0.05 and 3.7 +/- 0.2 vs. 3.4 +/- 0.2, P < 0.05). (4) Multivariate regression analysis showed that four variables was correlated independently with twin pregnancy including first treatment cycle of IVF-ET (OR = 1.82, P = 0.02), number of good quality embryos (OR = 1.35, P = 0.01), development stage score of the second best embryo (OR = 1.55, P = 0.009) and ovarian responsibility (OR = 0.96, P = 0.04). CONCLUSIONS: It is advisable to perform single embryo transfer. If patients are at high risk factors of twin pregnancy including initial IVF-ET treatment, good ovarian responsibility, more number of top-quality embryos and development stage score of the second best embryo transferred.

[Updated reproductive function of human sperm RNA].

The existence of RNA has been confirmed in human mature sperm, including mRNA and some members of the microRNA family. Different expressions of sperm mRNA have been found to be correlated with sperm motility and male reproduction. Some sperm specific mRNA and microNA play important roles in the regulation of sperm-oocyte fusion and early embryogenesis. Many published results indicate the variety of sperm RNA in composition and quantity as well as its indispensability for embryogenesis. Further researches on the function of sperm RNA will promote the progress in such fields as male infertility, human assisted reproduction technology and nuclear transfer.

[An analysis for the phenotype and genotype of autosomal dominant polycystic kidney disease from two Chinese families].

OBJECTIVE: To scan for mutations of polycystic kidney disease 1 gene (PKD1) in Chinese population in order to find some features about Chinese patients and a better approach to detect mutations. METHODS: Twenty-five PKD-affected individuals from twenty-one unrelated genealogies and sixteen controls participated in the study. Thirty-five blood samples and six tissues were obtained after receiving informed consent and were in accordance with institutional ethical guidelines. Genomic DNA was isolated from peripheral blood using standard procedures. PCR amplification of genomic DNA was performed to generate the aimed fragments. Amplified fragments were analyzed by denaturing gradient gel electrophoresis (DGGE). A GC clamp was attached to the 5' primer. After that, the abnormal fragments were sequenced on freshly amplified specific PCR products with the dideoxynucleotide chain termination method. Sequencing was performed for all samples to evaluate DGGE. RESULTS: Aimed fragments of exons 44 and 45 were amplified. DGGE detected eleven abnormal PCR fragments. Two novel mutations were identified by sequencing, included one nonsense mutation (C12217T) and one frameshift (12431delCT). In addition, one polymorphism (A50747C) was identified. The mutation detection rate is 8% in our study. CONCLUSION: Two novel pathogenic mutations were detected, including one nonsense mutation (C12217T) and one frameshift (12431delCT).