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Abstract Permanent preservation of genetic resources may be indispensable for the future of humanity. This requires liquid nitrogen, as is the case for preserving animal sperm. However, this technique is expensive and poses a risk of irrecoverable sample loss on non-replenishment of liquid nitrogen in case of natural disasters. In this study, we demonstrate that lyophilization may be used as a reliable method for long-term preservation of mouse sperm at room temperature. Sperm from four mouse strains were freeze-dried and stored in a non-temperature controlled room for 5–6 years. Although the ability of the stored sperm to activate oocytes had diminished slightly, healthy offspring were obtained by artificially activating the oocytes after sperm injection. Moreover, the birth rate did not decrease even after ≤ 6 years of storage. Furthermore, owing to its low cost, safety, and ease of storage at any location, we believe that this method could be a major mode of preserving mammalian genetic resources in the future.

Abstract Background In freeze–thawed embryo transfer (FET) cycles, hormone replacement treatment (HRT) is crucial for implantation and pregnancy maintenance. HRT typically continues until the 10th week of pregnancy owing to a luteoplacental shift, although a definitive HRT regimen remains undetermined. We present the case of a woman who underwent FET during an HRT cycle and ceased HRT after a negative pregnancy test at 3 weeks and 5 days, who went on to deliver a healthy baby. Case presentation A 30-year-old Japanese woman with primary infertility was scheduled for FET. Estrogen supplementation was initiated as part of the HRT cycle for endometrial preparation. After achieving an endometrial thickness of 8 mm, progesterone supplementation was commenced, and the transfer of a 4BB blastocyst occurred 5 days after initiating progesterone treatment. At a gestational age of 3 weeks and 5 days, her serum human chorionic gonadotropin (hCG) level was only 8.3 mIU/mL, leading to discontinuation of HRT due to the absence of pregnancy. However, at 6 weeks and 1 day, her serum hCG levels significantly rose to 9359 mIU/mL, prompting the resumption of HRT. Ultrasonography confirmed the presence of a gestational sac and cardiac activity in the uterus, and HRT was continued until the 10th week. Ultimately, she delivered a healthy female neonate vaginally, weighing 2601 g at 40 weeks and 6 days. Conclusions Progesterone supplementation is customary in FET with HRT cycles, although it has raised the possibility that there is demonstrating the potential for an ongoing pregnancy and resulting in a healthy baby under no progesterone replacement prior to the luteoplacental shift in this case. The duration and dosage of progesterone in luteal support for FET with HRT warrant further investigation.

Objective: To determine whether endometrial thickening is an important factor for pregnancy outcomes in platelet-rich plasma (PRP) treatment for patients with thin endometrium. Methods: Data from the registry database of the Japan Gynecologic PRP Study Group from April 2019 to October 2021 were analyzed retrospectively. A total of 208 patients who underwent single blastocyst transfer (SBT) after PRP due to thin endometrium (¡8 mm) in their previous cycle were included in the study. Their pregnancy outcomes were compared with their own historical data before PRP infusion. The same patient group was then divided into 136 patients whose endometrium thickened after PRP and 72 patients whose endometrium did not, and the pregnancy outcomes of the two groups were compared. Furthermore, to eliminate embryonic confounding, 28 patients who had single euploid blastocyst transfer (SEBT) with preimplantation genetic testing for aneuploidy (PGT-A) were selected from the same patient group and divided into two groups of 22 patients whose endometrium thickened and 6 patients whose endometrium did not, and the pregnancy outcomes were compared in the same method. Results: After PRP administration, the clinical pregnancy rate, live birth rate, and miscarriage rate were all significantly improved compared with the historical controls (34.1 vs 20.0, [Formula: see text]; 22.6 vs 3.9, [Formula: see text]; 33.8 vs 73.4, [Formula: see text]). There were no statistically significant differences in the respective rates between the “thickened endometrium” group and the “unthickened endometrium” group in SBT group (36.0 vs [Formula: see text], [Formula: see text]; 25.7 vs [Formula: see text], [Formula: see text]; 28.5 vs [Formula: see text], [Formula: see text], respectively). Likewise, in the SEBT with PGT-A group, there were no statistically significant differences in results between the two groups (36.4 vs [Formula: see text], [Formula: see text]; 36.4 vs [Formula: see text], [Formula: see text]; 0 vs [Formula: see text], respectively). Conclusions: PRP administration to patients with thin endometrium improves pregnancy outcomes even when endometrial thickening is not achieved.

Abstract Background Endometrial receptivity array (ERA) is used to determine the timing of embryo transfer (ET) synchronized with the window of implantation (WOI). The effectiveness and evaluation of ERAs in women with recurrent implantation failure remain controversial. We report the case of a patient with recurrent implantation failure that raises the issue of reproducibility of ERA tests. Case report A 36-year-old Japanese woman with secondary infertility who had previously given birth failed to conceive after three frozen-thawed embryo transfer (FET) cycles. An ERA test was conducted to confirm the WOI. The first ERA test was performed 125 h after progesterone exposure. The laboratory reported that the endometrium was in a non-receptive (post-receptive) phase, and recommended retesting 101 h after progesterone exposure. A simultaneous chronic endometritis (CE) test showed a score of 3. After the antibiotics administration to treat CE, the second ERA test was performed after 101 h of progesterone exposure. The laboratory reported that the endometrium had not reached the WOI and estimated the WOI to be 113 ± 3 h after progesterone exposure. The third ERA test was performed 113 h after progesterone exposure. The laboratory reported that the endometrium was in a non-receptive (pre-receptive) phase and estimated the WOI to be 137 ± 3 h after progesterone exposure. A CE test performed at the same time as the second and third ERA tests showed a score of 1 for the collected endometrium. According to the third ERA test results, the vitrified-warmed blastocyst was transferred at 137 h of progesterone exposure. Pregnancy was achieved and the patient had an uncomplicated vaginal delivery at 39 weeks. One year later, another pregnancy was achieved after FET at 137 h of progesterone exposure, and the patient delivered at 33 weeks due to an unexpected membrane rupture. Conclusion Because the results of the ERA test may vary in the presence of CE, CE should be diagnosed simultaneously with or before conducting ERA tests. If CE is diagnosed, ERA testing should be performed after treatment with antimicrobials or other drugs.

Endometrial receptivity array (ERA)—an objective tool used in assisted reproductive technology—is used for personalized embryo transfer in in vitro fertilization. Hydrosalpinx affects implantation through various mechanisms. However, its effects on ERA are not well established. In this case report, we present the diagnosis and treatment of a 34-year-old nulligravida woman with infertility for two years, obesity, double uterus with unilateral hydrosalpinx and right kidney deficiency. Based on ERA results, endometrial microbiome metagenomic analysis (EMMA), analysis of infectious chronic endometritis (ALICE), and CD138 immunostaining, the patient was treated with hormonal replacement cycle and amoxicillin/clavulanic acid. After one week of amoxicillin/clavulanic acid administration, the vitirified-warmed 4AA blastocyst was transferred to the left uterus—which was absent of hydrosalpinx and easily accessible to transfer and pregnancy was achieved. To the best of our knowledge, this case study is the first one in which we found that there were no differences between the left and right uterus in ERA, EMMA, ALICE, and CD138 immunostainings. Hence, we suggest that hydrosalpinx does not necessarily cause endometrial changes in all cases. Further research to evaluate the effects of hydrosalpinx on implantation with ERA and EMMA/ALICE is warranted.

Background and Aims: Little information is available for investigating the effect of the direction of sperm injected into the cytoplasm on clinical results for human oocytes following intracytoplasmic sperm injection (ICSI). This study aimed to examine the impact of the direction of sperm (head-first or tail-first) injected into the cytoplasm on fertilization and embryo development for high-elasticity or low-elasticity human oocytes following the Piezo-ICSI. Methods: We retrospectively investigated 2717 high-elasticity oocytes and 907 low-elasticity oocytes between May 2016 and December 2022. Of 2717 high-elasticity oocytes, 1148 were injected sperm head-first, and 1569 were injected sperm tail-first into the cytoplasm by Piezo-ICSI. Of 907 low-elasticity oocytes, 480 were injected head-first, and 427 were injected sperm tail-first into the cytoplasm. The membrane of high-elasticity oocytes was robust and was ruptured artificially by the piezo-pulse. The membrane of low-elasticity oocytes was weak and spontaneously ruptured without the piezo-pulse. The rates of degeneration, fertilization (2PN), 0PN, 1PN, [Formula: see text]3PN, abnormal cleavage, and good-quality day-3 embryos were evaluated in both groups. Results: Table 1 shows the effect of sperm direction on fertilization and embryo development for high-elasticity oocytes. The fertilization rate of tail-first group was significantly higher than the head-first group. The 0PN rate of tail-first group was significantly lower than the head-first group. The good quality Day-3 embryo rate of tail-first group was significantly higher than the head-first group. Table 2 shows the effect of sperm direction on fertilization and embryo development for low-elasticity oocytes. The fertilization rate of the tail-first group was significantly higher than the head-first group. However, no significant difference was observed in other comparable items. Conclusions: Tail-first sperm injection significantly improved the fertilization rates more than the head-first sperm injection regardless of the elasticity of the oocyte membrane following Piezo-ICSI.

Abstract Background Jejunogastric intussusception (JGI) is a rare, but potentially fatal complication that can occur following gastric surgery, and the reported incidence of JGI is as low as 0.1%. Early diagnosis and treatment are critical for JGI to prevent major complications such as bowel necrosis and death. Although emergency surgery is the standard treatment, endoscopic reduction has also been reported to be effective in JGI patients without bowel necrosis. Several early recurrent cases treated with surgical or endoscopic reduction have been reported. We report an extremely rare case of JGI after pancreaticoduodenectomy (PD) using Child’s procedure that was successfully treated with surgical reduction and fixation. Case presentation An 81-year-old man who had undergone PD using Child’s procedure 3 years ago presented to our hospital with epigastric pain and nausea. His vital signs were stable, and abdominal examination revealed mild tenderness with a palpable mass in the mid-epigastrium. Abdominal computed tomography (CT) and gastroscopy revealed a JGI of the efferent loop, and exploratory laparotomy was immediately performed. During the operation, the efferent loop showed no adhesions and was intussuscepted through the gastrojejunostomy into the gastric lumen. An incision in the anterior wall of the stomach revealed no evidence of ischemia of the intussusceptum. The efferent loop was reduced using Hutchinson’s maneuver and fixed to the afferent loop to prevent a recurrence. The postoperative course was uneventful, and there was no sign of recurrence 12 months postoperatively. Conclusions JGI after PD is an extremely rare, but has severe complications. Surgery might be the optimal treatment for JGI in terms of preventing recurrence, even in cases without bowel necrosis.

Background: The objective of this study was to investigate accurately the effect of the meiotic spindle imaging of human oocytes on their fertilization and embryo development by using the Piezo-ICSI. Methods: We retrospectively assessed 529 oocytes with the first polar body retrieved from 124 infertile couples (147 cycles) who attended our clinic between May 2016 and December 2018. Of these, 489 oocytes (92.4%) with visible meiotic spindle comprised the Spindle (+) group and 40 oocytes (7.6%) not observed meiotic spindle comprised the Spindle (−) group. The meiotic spindle was imaged using polarized light microscopy. The rates of fertilization, high-quality day-3 embryo, day-5 blastocyst, and high-quality day-5 blastocyst were evaluated in both groups. Results: The fertilization rates of Spindle (+) and Spindle (−) were 92.0% (450/489) and 70.0% (28/40), respectively; high-quality day-3 embryo rates were 62.9% (283/450) and 35.7% (10/28); day-5 blastocyst rates were 53.7% (205/382) and 32.1% (9/28); high-quality day-5 blastocyst rates were 29.8% (114/382) and 3.6% (1/28). Fertilization, high-quality day-3 embryo, day-5 blastocyst, and high-quality day-5 blastocyst rates were significantly higher in the Spindle (+) group than in the Spindle (−) group. Conclusion: Spindle imaging (i.e., oocytes with a visible meiotic spindle or with not observed meiotic spindle) influences the outcome of Piezo-ICSI in human oocytes, including fertilization and embryo development. To our knowledge, this study is the first to evaluate the effect of meiotic spindle imaging on fertilization or embryo development among oocytes after Piezo-ICSI. The meiotic spindle imaging could be the indicator for the quality management of medical doctors and embryologists.

Purpose: Male infertility is partially caused by an inappropriate lifestyle and comorbidities. In this study, we analyzed the prevalence of these factors and the effects of lifestyle modifications as part of male preconception care. Methods: Four hundred and two male partners of couples seeking conception with abnormal parameters upon the first semen analysis were enrolled. They were advised to modify their inappropriate lifestyle as male preconception care. Afterward, their general and male reproductive health was examined. Semen quality was compared before and after the promotion. Results: Smoking, chronic alcohol use, and genital heat stress were found in 22.6%, 47.0%, and 75.1% of patients, respectively. Palpable varicoceles, hypogonadism, obesity (body mass index ≧30 kg/m2), hypertension, zinc deficiency, hyperlipidemia, liver dysfunction, and diabetes mellitus were found in 25.9%, 17.0%, 7.0%, 14.9%, 16.2%, 37.0%, 26.9% and 3.4% of the participants, respectively; 98.8% of the patients had at least one factor. After the promotion, semen parameters and sperm DNA fragmentation were improved significantly. Improvement was found in those with palpable varicocele or hypogonadism but not in those with night work shift, abstinence (>3 days), erectile dysfunction, hypertension, obesity, zinc deficiency, or diabetes mellitus. Conclusions: Comorbidities and inappropriate lifestyle choices were common among men with infertility. The promotion of lifestyle modifications as part of male preconception care could improve semen quality without urologic intervention.

Abstract Background Subtotal cholecystectomy is an effective surgical method to decrease the risk of complications for gallbladders that are difficult to remove. However, there is a risk for postoperative refractory bile leakage through the gallbladder stump. Here, we report a new management technique involving the use of argon plasma coagulation (APC) to stop bile leakage after a subtotal cholecystectomy. Case presentation A 74-year-old man was referred to our hospital for abdominal pain and fever. Contrast-enhanced computed tomography of the abdomen showed fluid collection, such as an abscess, surrounding the gallbladder and hepatic flexure colon. The patient was diagnosed with colonic perforative peritonitis, and he underwent emergency surgery. On laparotomy, the abscess was observed outside of the hepatic flexure colon and gallbladder necrosis was detected. The neck of the gallbladder and the area close to the hepatoduodenal ligament was severely inflamed prohibiting dissection. The hepatic flexure colon was part of the abscess wall, and resection was needed. A subtotal cholecystectomy and right hemicolectomy confirmed peritonitis caused by cholecystic perforation. The mucous membrane of the gallbladder neck that remained was necrotic or detached. Therefore, the stump of the gallbladder was closed by primary sutures without cauterization of the mucosa. On postoperative day 6, bile leakage from the gallbladder stump was revealed. Percutaneous and endoscopic retrograde cholangiography drainage were performed. However, the liquid, which seemed to be secreted from the mucosa of the remnant gallbladder, was continuously obtained. We used APC to cauterize the gallbladder mucosa through the fistula of the abdominal drainage tube. Bile leakage and mucus discharge were improved after three rounds of APC cauterization. Conclusions APC effectively treated refractory bile leakage from a gallbladder stump after subtotal cholecystectomy for severe cholecystitis.

Background: Empty follicle syndrome (EFS) is a rare complication in which no oocytes are retrieved in oocyte pick-up (OPU) despite adequate controlled ovarian hyperstimulation (COH). Various studies and systematic reviews have reported that EFS is mainly caused by diminished ovarian reserve (DOR) because EFS tends to occur in patients with a poor response to COH. However, these factors do not explain all cases. Current knowledge of these pathological factors is limited, and treatment is unknown. This study aimed to find out the clinical predictors of EFS before OPU. Methods: In this study, 2342 cycles of 1148 cases that underwent OPU between January 2015 and November 2020 in two reproductive clinics were retrospectively enrolled. Ninety-one and 2251 cycles were classified as EFS (no cultivatable oocytes retrieved) and non-EFS (cultivatable oocytes retrieved), respectively. Results: The EFS and non-EFS incidence was 3.9% and 96.1%, respectively. The mean patient age in the EFS group was higher than that in the non-EFS group (40.3 ± 3.4 years vs. 37.9 ± 4.5 years, p < 0.001). Body mass indexes in the EFS and non-EFS groups were similar (21.7 ± 3.3 kg/m2 vs. 22.0 ± 3.5 kg/m2, p = 0.52). The anti-Müllerian hormone (AMH) levels, antral follicle count (AFC), and the number of follicles were lower in the EFS group than in the non-EFS group (1.0 ± 1.2 ng/mL vs. 2.7 ± 2.5 ng/mL, p < 0.001; 2.9 ± 2.2 vs. 8.5 ± 6.3, p < 0.001; 1.9 ± 1.4 vs. 6.0 ± 4.1, p < 0.001, respectively). However, unlike univariate analysis, logistic regression analysis showed no significant differences in age and AMH levels between the groups. The AFC and number of follicles were still significantly lower in the EFS than in the non-EFS group in multivariate analysis (odds ratio (OR), 1.301; 95% confidence interval (CI), 1.138–1.503; p < 0.05 and OR, 1.832; 95% CI, 1.488–2.3; p < 0.05, respectively). The EFS incidence rates for each follicle number just before OPU were 21.2%, 7.8%, 2.7%, and 1.2% with one, two, three, and four follicles, respectively. Not a single case of EFS was found among 410 cases with 10 or more follicles. Conclusions: The AFC and the number of follicles may be predictive of EFS.

Abstract Background The safety and feasibility of laparoscopic colectomy for T4 colorectal cancer remain controversial. We believe that setting a “Goal” that will guide the surgeons in returning from the deep layer could be the key to safe en bloc resection of neighboring organs. For descending colon cancer, the cranial-first approach makes it possible to clearly visualize the pancreas and origin of the transverse mesocolon, leading to safe splenic flexure mobilization and complete mesocolic excision, which is the strongest advantage of this approach. Case presentation A 75-year-old woman was diagnosed with T4 descending colon cancer invading the Gerota’s fascia. We performed laparoscopic left colectomy using the cranial-first approach to set a “Goal” at the inferior border of the pancreas for safe resection of the Gerota’s fascia. The total operative time was 233 min, and the estimated blood loss was 98 ml. She was discharged after surgery without postoperative complications. Pathological findings revealed the invasion into the Gerota’s fascia, and the resection margin was negative for cancer. Conclusions The cranial-first approach of laparoscopic left colectomy appears to be safe and feasible and could be a promising method for selected patients with T4 descending colon cancer invading the Gerota’s fascia.

Abstract Purpose Fertility preservation is an important issue for young cancer patients. Random‐start controlled ovarian stimulation and double ovarian stimulation have been proposed for efficient oocyte retrieval within the limited time before cancer therapy. We aimed to clarify the efficacy of these new protocols within the Japanese population. Methods We performed a retrospective observational study at a multicenter from February 2012 to August 2017. The study entailed 50 cycles with 34 patients who underwent fertility preservation due to breast cancer. Follicular phase or luteal phase ovarian stimulation with aromatase inhibitor was performed. A second ovarian stimulation was started with or without waiting until the next menstruation. We measured the number of retrieved oocytes and cryopreserved oocytes/embryos, the ratio of mature oocytes, and the fertilization rate. Results The numbers of retrieved oocytes and frozen oocytes/embryos were not significantly different between follicular phase and luteal phase ovarian stimulation. The number of retrieved oocytes was not reduced at the second ovum pick up compared to the first ovum pick up in the double ovarian stimulation. Conclusions Random‐start controlled ovarian stimulation and double ovarian stimulation with aromatase inhibitor for breast cancer patients were effective protocols for retrieving a greater number of oocytes within the limited time.

Abstract The epigenetic status of the genome changes dynamically from fertilization to implantation. In addition, the physiological environment during the process of gametogenesis, including parental age, may affect the epigenome of the embryo after fertilization. It is important to clarify the influence of parental age on gene expression in the embryo in terms of transgenerational epigenetics to improve the techniques currently used in assisted reproductive medicine. Here, we performed single-embryo RNA-seq analysis on human blastocysts fertilized by intracytoplasmic sperm injection, including from relatively elderly mothers, to elucidate the effects of parental age on the embryonic gene expression profile. We identified a number of genes in which the expression levels were decreased with increasing maternal age. Among these genes, several are considered to be important for meiotic chromosomal segregation, such as PTTG1, AURKC, SMC1B and MEIKIN. Furthermore, the expression levels of certain genes critical for autophagy and embryonic growth, specifically GABARAPL1 and GABARAPL3, were negatively correlated with advanced paternal age. In addition, levels of transcripts derived from major satellite repeats also decreased as the maternal age increased. These results suggest that epigenetic modifications of the oocyte genome may change with parental age and be transmitted to the next generation.

The feasibility and safety of laparoscopic surgery for locally advanced colorectal cancer remain controversial due to the high rate of incomplete resection and conversion to open surgery. Especially for T4 colorectal cancer, laparoscopic techniques are still demanding mainly because of the difficulty in distinguishing between inflammation and tumor involvement, which often lead surgeons to do overtreatment in surgery. We believe laparoscopic magnified and multidirectional approach might be useful for pathologically complete resection and minimizing an unnecessary extended surgery for these cases. A 49-year-old man was diagnosed with locally advanced T4 sigmoid colon cancer invading the urinary bladder and ureter. We performed laparoscopic anterior resection with en bloc resection of the urinary bladder and the left ureter. Total operative time was 462 min, and the estimated blood loss was 50 ml. This patient was discharged on the 28th day after surgery without any ostomies and urinary functional disorders. The magnified view by laparoscopic techniques from multiple directions would enable surgeons to set surgical landmarks for another approach, which is the key for safe and feasible laparoscopic surgery in patients with locally advanced T4 colorectal cancer.