Artificial Cycle with or without a Depot Gonadotropin-releasing Hormone Agonist for Frozen-thawed Embryo Transfer： An Assessment of Infertility Type that Is Most Suitable

The clinical outcomes of five groups of infertility patients receiving frozen- thawed, cleavage-stage embryo transfers with exogenous hormone protocols with or without a depot gonadotropin-releasing hormone （GnRH） agonist were assessed. A retrospective cohort analysis was performed on 1003 cycles undergoing frozen-thawed, cleavage-stage embryo transfers from January 1, 2012 to June 31, 2015 in the Reproductive Medicine Center of Wuhan General Hospital of Guangzhou Military Region. Based on the infertility etiologies of the patients, the 1003 cycles were divided into five groups： tubal infertility, polycystic ovary syndrome （PCOS）, endometriosis, male infertility, and unexplained infertility. The main outcome was the live birth rate. Two groups were set up based on the intervention： group A was given a GnRH agonist with exogenous estrogen and progesterone, and group B （control group） was given exogenous estrogen and progesterone only. The results showed that the baseline serum hormone levels and basic characteristics of the patients were not significantly different between groups A and B. The live birth rates in groups A and B were 41.67% and 29.29%, respectively （P〈0.05）. The live birth rates in patients with PCOS in groups A and B were 56.25% and 30.61%, respectively （P〈0.05）. The clinical pregnancy, implantation and on-going pregnancy rates showed the same trends as the live birth rates between groups A and B. The ectopic pregnancy rate was significantly lower in group A than in group B. We concluded that the live birth rate was higher and other clinical outcomes were more satisfactory with GnRH agonist co- treatment than without GnRH agonist co-treatment for frozen-thawed embryo transfer. The GnRH agonist combined with exogenous estrogen and progesterone worked for all types of infertility tested, especially for women with PCOS.

Morphological and ultrastructural changes in rat ovarian tissues after cryopreservation

Objective: The aim of the study was to observe and compare the effects of cryopreservation and thawing meth- ods on rat ovarian tissues. Methods: Twenty 5-6 weeks old SPF-SD female rats were randomly divided into two groups, with ten rats in each group. Freshly isolated ovaries saved as a control (group 1: fresh ovaries) in formalin-fixed or vitrified immediately after dissection (group 2: vitrified ovaries). Ovaries in vitrified group were processed into thin slices then cryo- preserved, stored in liquid nitrogen for 21 days, rapidly thawed and grossly examined. All of the collected ovaries underwent hematoxylin and eosin-stained paraffin serial sections and observed the microscopic evaluation in vitrified ovaries. Results: Grossly the vitrified ovaries turned pale color and the size was same as before freeze. The vitrified ovarian tissue had normal anatomical structures of cortex and medulla under the microscope and had no difference with the fresh control ovarian tis- sue. The number and distribution of the follicles were similar with the fresh ovarian tissue, but had smaller size and the gap between oocyte and the surrounding granulosa cells was increased. Few ooctyes were in irregular appearance however the morphology of follicular cells did not give a different appearance as compared to the fresh control ovarian tissue. Conclusion: Cryopreservation of ovarian tissues by vitrification method has some detrimental effect on the morphology of follicles but does not induce negative impact on the number, density and survival of the primordial ovarian follicles. However the whole follicle anatomical structures also had no significant changes.

Fertility preservation in female cancer patients

Cancer is not rare in younger women of reproductive age therefore the preservation of fertility among them has become a significant concern due to aggressive cancer therapy they must undergo.Today different strategies for fertility preservation are available in patients at risk.However,many of these available techniques are still experimental and have limited clinical experience.Under ethical principle,better interdisciplinary cooperation between clinicians and patients,appropriate counseling and further research on cryopreservation and transplantation techniques may enhance their success to preserve fertility of young cancer victims.In this article,the existing and emerging fertility preserving strategies in young cancer female as well as their safety and ethical issues are discussed.

Morphology and survival of cryopreserved-thawed ovarian tissues after heterotopic autotransplantation

Objective： The aim of our study was to observe the survival and morphological changes of thawed ovarian tis- sues after heterotopic transplantation. Methods： Twenty SPF-SD female rats （5-6 weeks old） were equally randomized into the control group and experimental group. In control group, the freshly isolated ovaries were fixed in formalin. In experimental group, the freshly isolated ovaries were vitrified immediately and cut into thin slices. After stored in liquid nitrogen for 21 days, the tissues of experimental group were rapidly thawed and transplanted into back muscles of rats for 2 or 4 weeks, respectively. After that, all rats in experimental group were sacrificed and the ovarian tissues were collected and fixed in 4% formaldehyde solution. Then the ovarian tissues were stained with HE and observed under the light confocal microscope. Re- suits： With the naked eyes, there was no specific alteration except the size reduction with color changing. Under microscopy, we found normal cortex and medulla in the ovary, and the primordial follicles and follicles in various stages were observed in the cortex. The normal oocytes in ovarian tissues of experimental group were significant decreased than in the control group. Conclusion： The ovarian tissues survive well in experimental group and there is no significant difference in the proportion of follicles between different times （2 and 4 weeks） after grafting. Our results suggest that thawed ovarian tissues could survive after heterotopic transplantation into back muscles of rat models and maintain their morphology and function.

Effects of heterotopic autotransplantation of cryopreserved-thawed ovarian tissues on sex hormone level

Objective: The aim of our study was to measure and compare the serum hormone level of transplant group with blank control and castrated control groups after heterotopic autotransplantation of cryopreserved-thawed ovarian tissues into back muscles. Methods: A total of 40 SPF-SD female rats(5–6 week-old) were randomly divided into three groups: blank control group(group A), castration control group(group B) and transplant group(group C). Ovaries were removed by surgical procedure, then after cryopreservation and thawing procedures the ovarian tissues were implanted into the back muscles of mice in group C. After 4 weeks of ovarian tissues transplantation, all rats blood sampling were measured for E2, LH and FSH hormone levels by ELISA. Results: E2 level was significantly higher in group C and group A than group B [(38.98 ± 5.66) pg/mL,(8.14 ± 3.24) pg/mL; P < 0.05) and [(36.30 ± 6.90) pg/mL,(8.14 ± 3.24) pg/mL; P < 0.05)]. However, E2 level in group C and group A had no significant difference. FSH level in group B, group A and group C was(18.87 ± 2.54) nmol/L,(7.77 ± 0.87) nmol/L and(9.39 ± 2.12) nmol/L respectively. FSH level increased significantly in group B compared with group A, and the difference had statistical significance(P < 0.05). FSH level was slightly increased in group C compared with group A, and the difference was not statistically significant(P > 0.05), but compared with group B, FSH level was significantly reduced and being statistically significant(P < 0.05). Conclusion: Autotransplantation of cryopreserved-thawed ovarian tissue into back muscles can sustain follicular development and re-establish endogenous hormone production by restoring the factors such as angiogenesis and innervations at the graft site.

Study on the Estrous Cycle Regularity of Cryopreserved Rat Ovarian Tissues after Heterotopic Transplantation

Objective: To evaluate the re-initiation of ovarian function in cryopreserved ovarian grafts by means of vaginal smear of transplant rats. Methods: A total of 40 SPF-SD female rats (5 - 6 week-old) were randomly divided into three groups (blank control, castration control and transplant group). Ovaries were removed by surgical procedure then after cryopreservation and thawing procedures the ovarian tissues pushed inside the back muscles gap in transplant group. On the first PO day, vaginal smear collection was daily initiated. After 30 days, the PO day when the estrous cycle was re-initiated was considered for analysis as well as the estrous days and the number of estrous cycles. Results: Normal control group had a regular estrous cycle, while the transplant group had an estrous cycle disorder within first 2 weeks and later 2 weeks after the transplantation while the duration of diestrus cycle lasted longer. At the same time, the castration group had lost the normal estrous cycle, keeping continue in the stage of diestrus. Conclusion: In transplanted animals the re-initiated ovarian function can be predicted with alteration between estrous and diestrus phases with predominant estrous irregularity. Moreover, short autotransplanted graft duration needs time to perfuse by new blood vessels and hormone secretions, so could not directly affect its target organs to function properly.

Characteristics of abnormal menstrual cycle and polycystic ovary syndrome in community and hospital populations

Background Polycystic ovary syndrome （PCOS） is considered to be the most common endocrine disorder in women of reproductive age.The involvement of an abnormal menstrual cycle in the etiology of PCOS remains unclear.We aimed to analyze the characteristics of abnormal menstrual cycles and their association with PCOS in community and hospital patient populations.Methods Women with PCOS identified from 2111 permanent female residents in the community of Beijing and 506 outpatients obtained from the reproductive clinic of Peking University Third Hospital were recruited for this study,comprising the PCOS community group and the PCOS hospital group, respectively.Each group was further divided into four subgroups according to the length of menstrual cycles： 〈21 days; 21-34 days; 35-60 days; and 〉60 days.Women in each group were interviewed using a questionnaire to assess factors including age, age of menarche, menstrual cycle history, related family history, and modified Ferriman-Gallwey （mF-G） score.All women received transvaginal ultrasound scan and had fasting blood samples taken for endocrine evaluation.A two-tailed P value of 〈0.05 was considered significant.Results In the community population, the prevalence of abnormal menstrual cycle was 27.19% （574/2111）.The prevalence of PCOS in the community was 6.11% （129/2111） according to Rotterdam criteria.In the community group,the most common menstrual cycle length was 35-60 days, whereas for the hospital group, it was 〉60 days.In both the community and hospital groups, the most common phenotype of PCOS was that of Oligo/amenorrhea＋PCO＋hyperandrogenism （HA） （O＋P＋H） （P=0.000）.With increasing cycle length of 35-60 days to 〉60 days, the percentages of Oligo/amenorrhea＋PCO （O＋P） and O＋P＋H were found to significantly decrease in the community group and significantly increase in the hospital group （P=0.000 for each）.In the hospital group, as the menstrual cycle length increased from 35-60 days to 〉60 days, the rate of spontaneous abortion increased significantly （P=0.000）, meanwhile the rate of poorly-secreted endometrium and abnormal endometrial hyperplasia increased significantly （P=0.000）.Conclusions The prevalence of PCOS in the Beijing community of women was 6.11%.Oligo/arnenorrhea was the most common type of abnormal menstrual cycle and may be an indicator for PCOS and endometrial lesions.Gynecologists should seek relevant medical information from women in the community to promptly diagnose PCOS and then follow up patients for potential development of subsequent complications.

Clinical Analysis on Laparoscopic Salpingectomy for Ectopic Pregnancy Following IVF/ICSI-ET

Objective To investigate whether laparoscopic surgery is valuable during early pregnancy and its influence on delivery outcome of the patients.Methods A retrospective analysis was undertaken of 14 heterotopic pregnancies conceived by IVF/ICSI-ET and underwent laparoscopic salpingectomy from December 2001 to September 2009.Results The mean age of 14 patients was 29 ± 5.3 years with range of 21-39 years.The ectopic pregnancy was located in the ampulla（11 patients,78.57%） and isthmus（3 patients,21.43%）.All 14 patients underwent successful laparoscopic salpingectomy without complications.Eleven（78.57%） patients delivered live singleton infants at full term via caesarean section while 3（21.43%） had spontaneous abortion.The birth weight of viable newborns ranged from 2 600 g to 3 720 g and had a high Apgar score.Conclusion During early pregnancy,the heterotopic pregnancy should be diagnosed and treated early and laparoscopic salpingectomy can be a safe and effective choice.The well experienced surgeons and accessibility of the modern equipments are mandatory.

Relationship between hyperemesis gravidarum and Helicobacter pylori seropositivity

Early pregnancy is commonly characterized by the disturbance of digestive function, characteristically manifested by nausea and vomiting. These symptoms during early pregnancy usually disappear spontaneously after about 12 weeks. In a minority of pregnant women, these symptoms are very severe, and are associated with weight loss, dehydration, acidosis from starvation,

Strong association of SLCIA1 and DPF3 gene variants with idiopathic male infertility in Han Chinese

Male infertility is a multifactorial syndrome encompassing a wide variety of disorders. In recent years, several genome-wide single-nucleotide polymorphism （SNP） association studies （GWAS） have been performed on azoospermia and/or oligozoospermia in different populations including two GWAS on nonobstructive azoospermia in China; however, the association of SNPs with idiopathic male infertility, especially asthenozoospermia and oligozoospermia, and their correlation with semen parameters are still not clear. To investigate genetic variants associated with idiopathic male infertility （asthenozoospermia, oligozoospermia, and oligoasthenozoospermia） in Chinese Han people, 20 candidate SNPs were selected from GWAS results and genotyped using the Sequenom MassARRAY assay. A total of 136 subfertile men and 456 healthy fertile men were recruited, rs6476866 in SLCIA1 （P = 1.919E-4, OR = 0.5905, 95% Ch 0.447-0.78） and rs10129954 in DPF3 （P = 0.0023, OR = 2.199, 95% Ch 1.311-3.689） were strongly associated with idiopathic male infertility. In addition, positive associations were observed between asthenozoospermia and rs215702 in LSM5 （P= 0.0016, OR = 1.479, 95% CI： 1.075-2.033） and between oligoasthenozoospermia and rs2477686 in PEXIO （P= 0.0011, OR = 2.935, 95% Ch 1.492-5.775）. In addition, six SNPs （rs215702 in LSMS, rs6476866 in SLCIA1, rs10129954 in DPF3, rs1801133 in MTHFR, rs2477686 in PEXIO, and rs10841496 in PED3A） were significantly correlated with semen quality alterations. Our results suggest that idiopathic male infertility in different ethnic groups may share the same mechanism or pathway. Cohort expansion and further mechanistic studies on the role of genetic factors that influence spermatogenesis and sperm progressive motility are suggested.

Alternative splicing patterns of doublesex reveal a missing link between Nix and doublesex in the sex determination cascade of Aedes albopictus

Sexual development in insects is regulated by a complicated hierarchical cascade of sex determination.The primary signals are diverse,whereas the central nexus doublesex(dsx)gene is relatively conserved within the pathway.Aedes(Stegomyia)albopictus is an important vector with an extensive worldwide distribution.We previously reported that Ae.albopictus dsx(Aalbdsx)yields one male-(Aalbdsx^(M))and three female-specific isoforms(Aalbdsx^(F1-3));however,the spatiotemporal expression profiles and mechanisms regulating sex-specific alternative splicing require further investigation.In this study,we demonstrated that the Aalbdsx^(M) messenger RNA(mRNA)represents the default pattern when analyzed in human foreskin fibroblasts and HeLa cells.We combined reverse transcription polymerase chain reaction with RNA immunoprecipitation using specific antibodies against tagged Ae.albopictus male-determining factor AalNix and confirmed that AalNix indirectly regulates dsx pre-mRNA and regulates its alternative splicing.During the early embryo stage(0-2 and 4-8 h),maternal dsx^(F) and default splicing dsx^(M) were detected in both sexes;the expression of dsx^(M) then decreased until sufficient AalNix transcripts accumulated in male embryos at 20-24 h.These findings suggest that one or more potential dsx splicing enhancers can shift dsx^(M) to dsx^(F) in both sexes;however,the presence of Nix influences the function of this unknown splicing enhancer and ultimately leads to the formation of dsx^(M) in males.Finally,our results provide important insight into the regulatory mechanism of dsx alternative splicing in the mosquito.

Effect of Mitochondrial Transplantation from Cumulus Granular Cells to the Early Embryos of Aged Mice

Objective To assess the role of mitochondria in the early embryonic development of ageing mice.Methods Mitochondria isolated from cumulus granular cells of aged mice were microinjected into oocytes or zygotes of aged mice. In the setting of oocyte injection, mitochondria were transferred via intracytoplasmic sperm injection （ICSI＋MIT）, and ICSI without mitochondrial transfer. In the setting of zygote injection, mitochondria were directly microinjected into fertilized oocytes （MIT）, and those injected with buffer alone （mock injection） or not injected （uninjected） served as controls.Results Although the rates of oocyte cleavage between ICSI and ICSI＋MIT groups were not statistically different （P〉0.05）, the rate of blastocyst in the ICSI＋MIT group was significantly higher than that in ICSI group （P〈0.05）. Although both the cleavage and blastocyst rates of mock injection group were significantly lower than those of uninjected group （P〈0.05）, likely due to mechanical damages of the cells by microinjection, the decrease of these rates was prevented by mitochondrial transfer. After mitochondrial transfer, the rates of both cleavage and blastocyst were significantly improved over the mock-iniection group （P〈0.05）.Conclusion Mitochondrial transplantation can improve the developmental potential of early embryos of aged mice.

A Systemic Review and Meta-analysis of the Effect of SARS-CoV-2 Infection on Sperm Parameters

Objective.Several studies examined the putativefets of SARS-Cov-2infectionon spem parametersHoweve,the resuls remain controversial.In this study,we conducted the most up-to-date systematic review and meta-analysis to investigate the effect of SARS-CoV-2 infection on sperm quality in COVID-19-positive and COVID-19-negative male participants.Method.Seven databases were searched forliterature released through June 10,2022,containing estimates for the outcomes of interest Using a random-fects model(REM)or a fixed-ffects model(FEM),we analyzed the pooled results.The quality of all included studies was asssed by the Newcastle-Ottawa scale In addition,we performed aquanttative and subgroup analysis of semen data across all included studies.Results.Fourteen studies were extracted from 10 publications,involving a total of 1174 participates for meta-analysis.Sperm parameters of 521 COVID-19 male patients and 653 controls were analyzed.In 8 case-control studies,the pooled mean dfference(MD)of total sperm motility was-5.37%(95%confidence interval(CI):-8.47 to-2.28:<0.05),suggesting that total motility was significantly impaired inmale COVID-19 cases.Subgroup analysis showed a significant decreasein semen volume,sperm concentration,and total motility in 238 patients with arecovery timeof less than 90 days.Moreover,in the other 6 included pre-to post-COVID-19 studies,the pooled MDs of sperm concentration,total sperm count,total motility,progressivemotility,and normal morphology were6.54×10^(6)/ml(95%CI:-10.27 to-2.81;p<0.05),38.89×10^(6)(95%CI:-59.20 to-18.58;p<0.05),-7.21%(95%CI:-14.36 to-0.07;p<0.05),-5.12%(95%CI:-8.71 to-1.53;p<0.05),and-1.52%(95%CI:-2.88 to-0.16;p<0.05),respectively,which indicate SARS-CoV-2 infection significantly affected these five sperm parameters.Conclusion.Our results revealed that SARS-CoV-2 infection was significantly correlated with decreased sperm quality.Of six sperm parameters,total motility and sperm concentration were the most significantly decreased parameters.These results suggest a possible negative influence of SARS-CoV-2 infection on testicular function and male fertility.Given the potential detrimental effect of SARS-CoV-2 on semen quality,male reproductive health should be monitored closely in patients with COVID-19.This trial is registered with CRD42021275823.

Different Pretreatments before Application of GnRH Antagonist Protocol in Controlled Ovarian Hyperstimulation

Objective To analyze the clinical features and outcome of patients who used different pretreatments before application of gonadotropin-releasing hormone antagonist protocol during in vitro fertilization - embryo transfer （IVF-ET） cycles, and to explore how effective to use the antagonist protocol. Methods A retrospective analysis was performed. All the ET cycles were divided into three groups, group A （n=125） used short acting GnRH agonist before GnRH antagonist treatment, group B （n=113） used short-acting oral contraceptives before GnRH antagonist treatment, group C （n=81） was untreated before GnRH antagonist treatment. All the patients had no tubal fluid, endometrial polyps and no anatomical abnormalities of the uterus, from April 2010 to December 2010. The patient＇s age, dose and duration of gonadotropin （Gn） treatment, the serum LH and E2 levels on the day of hCG injection, the number of oocytes retrieved, the rates of good-quality embryos, the clinical pregnancy rates were compared. At the same time, 261 GnRH agonist long protocol cycles （group D） were selected at the same period as further comparison. Results The patients in group C （32.9 ~ 4.8 years） were significantly older than those in groups A and B （31.6 ___＋3.7 years, 31.2 ___%4.1 years）（P 〈0.05）. The dose and the duration of Gn in group C were significantly lower than those in groups A and B. The serum LH level on the day of hCG injection in group A and group B was significantly lower than that in group C （P 〈0.05）, especially in group A. The endometrium was the thinnest in group B. There were no significant differences in the fertilization rates and the good-quality embryosrates among them. The clinical pregnancy rate of group B decreased significantly compared with groups A and C （P〈0. 05）. The clinical pregnancy rate of group C was the highest among them. There was no significant difference of clinical pregnancy rates between group C and group D （37% vs 40.2%,P〉0.05）. However, the dose （19.8 ±6.6 ampoule vs 26.4 ±8.1 ampoule） and the duration （9.0± 1.6 d vs 11.6±2.5 d） of Gn treatment in group C were decreased significantly than those in group D, P〈0.05. Conclusion The short acting GnRH agonist used before GnRH antagonist treatment during IVF-ET cycles failed to improve the pregnancy rates, the use of short-acting oral contraceptives before GnRH antagonist treatment makes the pregnancy rates decrease significantly, but untreated before GnRH antagonist protocol can get a better clinical outcome compared with agonist long protocol Untreated GnRH anagonist protocol is the best GnRH anagonist protocol.