nishizawa haruki

BACKGROUND: LHX8 gene encodes a germ cell specific transcription factor that is required for oocyte development. We evaluated two unrelated women with primary infertility who showed reproducible oocyte abnormalities across in vitro fertilization cycles, and we performed genomic and functional assays to clarify the role of LHX8. RESULTS: Whole exome sequencing identified heterozygous loss-of-function variants in LHX8 (NM_001001933.1) in both patients: c.778 C > T (p.Gln260Ter) in family 1 and c.581-1G > A in family 2. Both variants met the American College of Medical Genetics and Genomics criteria for likely pathogenicity. The two patients had high proportions of degenerated or immature oocytes and showed consistent morphologic features, including multiple cytoplasmic vacuoles, impaired zona pellucida function with accumulation of sperm in the perivitelline space, and poor embryo development. The splice site variant was inherited from a fertile mother, which indicates incomplete penetrance. A minigene assay confirmed the use of a cryptic acceptor site that produced a one nucleotide deletion and a frameshift, consistent with loss of function. CONCLUSIONS: These findings expand the phenotypic spectrum of LHX8 related infertility and provide mechanistic evidence that partial reduction of LHX8 activity compromises oocyte quality. Recognition of the characteristic morphology may guide genetic testing and counseling in cases of unexplained infertility. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13048-026-01978-2.

OBJECTIVE: Myotonic dystrophy type 1 (DM1) is an autosomal dominant neurodevelopmental disorder caused by CTG repeat expansion in the DMPK gene. Although the clinical classification of DM1 is determined by the CTG repeat length in DMPK, conventional sizing relies on Southern blotting, which is a suboptimal method in prenatal and PGD contexts as it requires large amounts of genomic DNA. We here evaluated the utility of nanopore long read sequencing (LRS) for DM1 diagnosis in these contexts. METHOD: LRS was performed with adaptive sampling or CRISPR/Cas9-mediated enrichment targeting DMPK. The use of whole genome amplified DNA (WGA-DNA) prepared with RepliG was also assessed. RESULTS: Adaptive sampling and Cas9-based LRS enabled detection of both the normal and expanded alleles. Further, LRS with CRISPR/Cas9-mediated enrichment improved efficiency and enabled accurate sizing of expanded CTG repeats exceeding 1000 units. In contrast, the use of whole genome amplified DNA prepared with RepliG did not permit reliable CTG repeat sizing, even when combined with adaptive sampling or CRISPR/Cas9. CONCLUSION: Nanopore sequencing can potentially replace Southern blotting for prenatal DM1 diagnosis, including repeat sizing. However, further improvement is needed for PGD using WGA-DNA.

Renal cell carcinoma is a malignant tumor that is prone to distant metastasis, primarily developing in the lungs, bones, lymph nodes, liver, and brain. Conversely, ovarian metastasis is rare, and its clinical characteristics and optimal treatment strategies remain unestablished. We report two cases with renal cell carcinoma and concomitant ovarian metastasis. Case 1 was of a 52-year-old woman with a metastatic tumor in the left ovary detected 9 years after radical nephrectomy for left-sided clear cell renal cell carcinoma. Case 2 was of a 56-year-old woman with a metastatic tumor in the right ovary detected 15 years after radical nephrectomy for right-sided clear cell renal cell carcinoma. In both cases, preoperative imaging revealed a pelvic tumor with a strong contrast effect. Surgery was performed for diagnosis and treatment of suspected ovarian metastasis from the renal cell carcinoma. Rapid intraoperative diagnosis was performed to determine the surgical approach, and the diagnosis of ovarian metastasis from renal cell carcinoma was made. Subsequently, bilateral adnexectomy was performed, and following pathological examination led to the definite diagnosis of ovarian metastasis from clear cell renal cell carcinoma. Long after radical nephrectomy, ovarian metastasis from the renal cell carcinoma can occur. Therefore, continuous follow-up is important. Complete resection of ovarian metastases from renal cell carcinoma may contribute to improved prognosis; however, more case studies are needed to establish standard treatment.

OBJECTIVES: Hormone replacement therapy (HRT) is considered for ovarian cancer patients who develop menopausal symptoms, dyslipidemia, and osteoporosis due to iatrogenic menopause caused by surgery or anticancer drug treatment. However, there have been few reports on HRT administration methods and those that evaluate cancer recurrence and complications of HRT. METHODS: We examined the administration method, adverse reactions and cancer recurrence in 28 patients who received HRT after ovarian cancer surgery at our hospital. RESULTS: All patients received estradiol monotherapy, and cancer recurrence was observed in four patients (14.3%); adverse reactions included skin eruption in two patients (7.1%), there were no other serious adverse reactions noted. CONCLUSION: The method and duration of HRT administration and the timing of HRT discontinuation remain debated. Thus, a large-scale survey and standardization of HRT administration methods after ovarian cancer surgery in Japan are needed.

RESEARCH QUESTION: What is the prevalence of tubulin beta 8 class VIII (TUBB8) variants in Japanese women with oocyte/zygote/embryo maturation arrest (OZEMA), and what are the phenotype-genotype correlations in this population? DESIGN: This multicentre retrospective study analysed 39 Japanese women with primary infertility characterized by OZEMA. Whole-exome sequencing was performed to identify TUBB8 variants, followed by Sanger sequencing, in-silico analysis, and structural modelling. Clinical data were collected across multiple treatment cycles, including oocyte maturation, fertilization, and embryonic development. Time-lapse imaging observed abnormal fertilization processes. RESULTS: Six TUBB8 variants were identified in 15.4% (6/39) of participants, including one novel homozygous nonsense variant (p.Thr136*) and two novel heterozygous missense variants (p.Phe294Leu and p.Trp344Arg). All six patients exhibited degrees of oocyte maturation arrest, predominantly at metaphase I. The novel homozygous nonsense variant was uniquely associated with multipronuclear (MPN) zygote formation. Both novel heterozygous missense variants, one confirmed de novo, caused complete metaphase I arrest. Time-lapse imaging in the patient with the homozygous nonsense variant demonstrated impaired polar body extrusion characterized by unusually broad cytoplasmic protrusions. These protrusions were reabsorbed into the oocyte, resulting in MPN zygotes. CONCLUSIONS: In this first study of TUBB8 in Japanese patients with OZEMA, three novel variants with unique phenotype-genotype correlations were identified. The phenotypes observed with the homozygous p.Thr136* variant suggest that complete TUBB8 loss disrupts not only meiotic progression but also post-fertilization events, including second polar body extrusion and pronuclear formation. These findings expand the spectrum of TUBB8-related reproductive phenotypes, and suggest that genetic testing can help avoid repeated unsuccessful treatments for primary infertility.

OBJECTIVE: Uterine carcinosarcoma (UCS) is a rare and aggressive gynecological cancer with high recurrence rates and is associated with a poor prognosis. It is also characterized by a high frequency of copy number alterations (CNAs). This study aimed to determine which gene CNA contributes to progression-free survival (PFS) in patients with UCS to identify potential prognostic biomarkers and therapeutic targets. METHODS: DNA was extracted from formalin-fixed paraffin-embedded tissues of surgical specimens from 24 patients with UCS who were treated at Fujita Health University. Using the PleSSision-Rapid test, the mutation information of 145 cancer genes was analyzed. Oncoplot analysis was used to visualize gene mutation profiles. χ2 test, Kaplan-Meier analysis, and log-rank test were used for statistical analyses. RESULTS: The most frequently observed gene mutation was TP53 in the 24 UCS cases studied, while genes associated with the PI3K/AKT/mTOR signaling pathway, including PIK3CA, PTEN, PIK3R1, and PIK3R2 were commonly detected. In the χ2 test analysis, TP53 loss (p=0.029), PIK3CA amplification (p=0.034), and TSC2 loss (p=0.034) were significantly associated with recurrence. Kaplan-Meier survival analysis demonstrated a significant association between PIK3CA amplification and TSC2 loss with PFS, as determined by the log-rank test (p<0.05). CONCLUSION: In patients with UCS, TSC2 loss is linked to poorer PFS, highlighting its utility as a prognostic marker. The association between TSC2 loss and increased recurrence risk highlights the potential therapeutic advantage of targeting the mTOR pathway in TSC2-deficient tumors.

INTRODUCTION: Alternative RNA splicing adds diverse variations to gene function, and its abnormalities are occasionally associated with the etiology of disease. We examined this possibility in pre-eclampsia. METHODS: We performed transcriptome analysis of placentas from pre-eclamptic and normotensive pregnancies and screened for disease-specific aberrant splicing. RESULTS: We identified aberrant splicing at exon 14 in the ZC3H4 gene. This in-frame exon is generally skipped in placentas from normal pregnancies but often observed in those from pre-eclampsia patients. The level of exon inclusion did not correlate with disease severity, such as blood pressure or fetal weight, but showed an association with the decrease in placental weight. Significantly, placental blood flow resistance measured by Doppler ultrasound correlated with the level of ZC3H4 exon 14 inclusion, suggesting that this retention leads to the onset and/or symptoms of pre-eclampsia. ZC3H4 is known to act on transcriptional regulation via suppression of lncRNA expression. Moreover, the SOD1 gene, encoding superoxide dismutase that eliminates toxic free superoxide radicals, was identified in the downstream gene group for ZC3H4. Indeed, the expression of SOD1 was found in this current study to be decreased in the pre-eclamptic placenta in correlation with the levels of ZC3H4 exon 14 retention. DISCUSSION: Aberrant splicing of ZC3H4 gene may induce excessive oxidative stress in the placenta via the downregulation of downstream SOD1 expression thereby leading to the onset and development of pre-eclampsia.

Introduction Cervical cancer is the fourth most common malignancy in women and is primarily caused by persistent infection with high-risk human papillomavirus (HPV). In addition, host immune responses, genetic factors, and lifestyle habits also have etiological roles. The cervicovaginal microbiome undergoes dynamic changes during menopause, which may be involved in the progression of cervical neoplasia. We aimed to elucidate the association between cervical microenvironmental changes and the progression of cervical neoplasia before and after menopause by integrating analyses of the cervical microbiome, related metabolites, cytokines, and microRNAs. Methods A total of 248 HPV-positive women with cervical neoplasia, including 17 with cervical intraepithelial neoplasia (CIN1), 80 with CIN2, 82 with CIN3, and 69 with squamous cell carcinoma (SCC), were enrolled. As normal controls, 48 HPV-negative healthy women were included. Each group was stratified based on the mean menopausal age of 50 years. Cervical mucus was analyzed according to the methods outlined below. The microbiota was profiled by 16S rRNA gene sequencing, metabolites were analyzed by ultra-HPLC-tandem mass spectrometry, RT-qPCR was used for miRNA expression analysis, and RANTES levels were quantified by multiplex bead array. Data analysis was performed using MicrobiomeAnalyst and MetaboAnalyst. Results In the SCC group, Prevotella and Atopobium were the key bacterial genera among the younger group, while Peptoniphilus, Fusobacterium, and Porphyromonas were more prevalent in elderly group (LDA score &amp;gt; 4.5). We observed a consistent positive correlation between Atopobium and xanthine in younger groups with CIN2 or worse (p &amp;lt; 0.0001). However, no such correlations were detected in elderly women. In addition, Atopobium, Adlercreutzia, and Gardnerella showed significant positive correlation with nicotinic acid in younger women with SCC compared to the elderly women (p &amp;lt; 0.0001). In the younger SCC women, several metabolites were significantly elevated in groups with high expression levels of RANTES, miR-20b-5p, and miR-155-5p. Conclusion The cervical microbiome undergoes changes during menopause, and may influence disease progression by interacting with metabolites, cytokines, and miRNAs. These results highlight the potential for personalized medicine for cervical cancer that is tailored to different age groups.

Objectives: We previously reported a high prevalence of hypovitaminosis D (25OHD < 20 ng/mL) in Japanese pregnant women with threatened premature delivery. This study aimed to assess nutritional status and its relationship with bone-related markers and microarchitecture, as measured using quantitative ultrasonography (QUS), in Japanese women during the perinatal period. Methods: We recruited Japanese women who had just delivered at Fujita Health University Hospital (n = 103, cesarean/vaginal delivery = 50/53, age 33.9 ± 4.9 years). On the third day postpartum, their calcaneal QUS was measured, and fasting blood samples were collected. Results: The mean total energy intake (1720 ± 298 kcal/day) was lower than the normal range for Japanese women (2100 kcal/day). Their calcium intake (446 ± 130 mg/day) was significantly below the recommended daily intake (RDI) in Japan (660 mg/day), with 95% of participants consuming less than the RDI. Although the average vitamin D intake (8.7 ± 1.8 μg/day) met the Japanese RDI (8.5 μg/day), 36% of participants consumed less than the RDI. Calcium intake was positively associated with the intake of lipids, protein, and vitamins A, D, and K. Additionally, calcium intake but not vitamin D intake tended to correlate with serum 25-hydroxyvitamin D (25OHD) levels. The QUS indices showed no significant association with calcium or vitamin D intake. Conclusions: During the perinatal period, Japanese women had low calcium intake and relatively low vitamin D intake, accompanied by reduced 25OHD levels. These findings highlight the need for public health recommendations and policies to promote adequate calcium and vitamin D intake during pregnancy.

BACKGROUND: When using assisted reproductive technology, there are cases where, despite the transfer of a good embryo, sometimes pregnancy may not be the case. Thus, during hormone replacement cycle implantation, it is important to synchronize the number of days of progesterone administration with the degree of embryo maturity. This study aimed to compare the outcomes of the administration of oral dydrogesterone for the duration of progestin use during the hormone replacement cycle for frozen-thawed blastocyst transfer. MATERIAL AND METHODS: The primary outcome of this study was the clinical pregnancy rate. We performed a retrospective cohort study of patients who underwent frozen-thawed blastocyst transfers between January 2017 and December 2024. According to our standard protocol, a vitrified-warmed blastocyst transfer was performed using dydrogesterone, which was administered orally at our center. A total of 554 cases were included in the study. Using the Gardner classification to evaluate the quality of blastocysts, grade AA was classified as the best quality, the AB/BA group as good quality, and the BB group as fair quality. We classified the 554 cases into 317 AA, 163 AB/BA, and 74 BB cases using the Gardner classification. Based on the duration of progestin administration, patients were divided into four groups: 120 hours (120 h), 132 hours (132 h), 144 hours (144 h), and 156 hours (156 h). We used the Shapiro-Wilk method and the Steel-Dwass test to determine whether there were differences in patients' background age and BMI among the four groups (120 h, 132 h, 144 h, and 156 h). We used Fisher's exact test and the Bonferroni method to determine whether there were differences in the final outcome of pregnancy rate between the four groups of 120 h, 132 h, 144 h, and 156 h. RESULTS: In the analysis of all embryos, the pregnancy rate at each timepoint of the primary evaluation was significantly higher in the 144-h group than in the 132-h group. Next, on analyzing the results by embryo grade, there was no difference in the pregnancy rate at each timepoint in the AA group. In the AB/BA group, the pregnancy rate was higher in the 144-h group than in the 132-h group. In the BB group, the pregnancy rate was higher in the 144-h group than in the 132-h group. CONCLUSION: This study clarified two aspects. First, the pregnancy rate in the 144-h group was significantly higher than that in the 132-h group in the analysis of all embryos. Second, the window of implantation may be more important for poor-quality embryos. This study showed that the oral administration of dydrogesterone requires a window of implantation of at least 144 hours.

Although organ transplantation is becoming general practice, little is known about the safety of delivery. This is the first known case that describes injury to the kidney by the uterine fundal pressure maneuver during cesarean section in a pancreas and kidney transplant recipient. A 40-year-old pregnant woman (gravida 0, para 0) was referred to our clinic. She had undergone living donor kidney transplantation 11 years earlier and brain-dead donor pancreas transplantation 1 year earlier owing to type 1 diabetes. Cesarean section was indicated when the patient's blood pressure was 150/100 mmHg at 37 weeks. We pushed the uterine fundus during delivery of the infant, with our usual caution. Serum creatinine levels were 1.6-2.6 mg/dl postoperatively. As this elevation was considered to be due to kidney graft dysfunction, we performed computed tomography, which revealed a hematoma around the kidney graft. Fifteen days after the cesarean section, surgical removal of the hematoma was performed by the transplant surgery team. Following hematoma removal, the serum creatinine level decreased to <1.4 mg/dl. We present a case of kidney graft injury during cesarean section in a pancreas and kidney transplant recipient.

OBJECTIVES: Chemotherapy-induced peripheral neuropathy (CIPN), a frequently occurring adverse event associated with paclitaxel/carboplatin (TC) combination therapy, causes limb pain and markedly reduces the patient's quality of life. Since zinc has been reported to be associated with neuropathic pain, we investigated the relationship between CIPN due to TC therapy and serum zinc levels. METHODS: The study included 13 patients with gynecological cancer whose serum zinc levels were measured before and during TC therapy. CIPN was classified into severity grades based on the Common Terminology Criteria for Adverse Events v5.0. A retrospective analysis was conducted on the relationship between the serum zinc level before TC therapy (PreZn), the minimum serum zinc level measured during TC therapy (MinZn), the MinZn/PreZn ratio, the number of TC treatment cycles, and the maximum grade of CIPN (MaxG) using Pearson's correlation coefficient. Moreover, an analysis was also conducted on clinical factors influencing MaxG, as well as fluctuations in serum zinc levels and CIPN grades for each cycle of TC therapy. RESULTS: A negative correlation was observed between the MinZn/PreZn ratio and MaxG (r=-0.557, p=0.048). The clinical factors influencing CIPN remained unclear, and the decrease in serum zinc levels and the aggravation of CIPN plateaued after the third cycle. CONCLUSIONS: If a decrease in serum zinc levels during TC therapy is smaller than before therapy, it may imply the existence of a causal relationship that suppresses the aggravation of CIPN.

Cytogenetic information about the product of conception (POC) is important to determine the presence of recurrent chromosomal abnormalities that are an indication for preimplantation genetic testing for aneuploidy or structural rearrangements. Although microscopic examination by G-staining has long been used for such an evaluation, detection failures are relatively common with this method, due to cell-culture-related issues. The utility of low-coverage whole-genome sequencing (lcWGS) using short-read next-generation sequencing (NGS) has been highlighted recently as an alternative cytogenomic approach for POC analysis. We, here, performed comparative analysis of two NGS-based protocols for this purpose based on different short-read sequencers (the Illumina VeriSeq system using a MiSeq sequencer and the Thermo Fisher ReproSeq system using an Ion S5 sequencer). The cytogenomic diagnosis obtained with each NGS method was equivalent in each of 20 POC samples analyzed. Notably, X chromosome sequence reads were reduced in some female samples with both systems. The possibility of low-level mosaicism for monosomy X as an explanation for this was excluded by FISH analysis. Additional data from samples with various degrees of X chromosome aneuploidy suggested that it was a technical artifact related to X chromosome inactivation. Indeed, subsequent nanopore sequencing indicated that the DNA in the samples showing the artifact was predominantly unmethylated. Our current findings indicate that although X chromosome data must be interpreted with caution, both the systems we tested for NGS-based lcWGS are useful alternatives for the karyotyping of POC samples.

INTRODUCTION: The da Vinci SP surgical system is a surgical platform capable of implementing robotic-assisted surgery through a single port and was first introduced in Japan at our hospital. In this paper, we describe our experience of the initial introduction of the da Vinci SP surgical system and its surgical outcomes. This is the first report on the surgical outcomes of using da Vinci SP, and its comparison with the conventional system in Japan. METHODS: After developing an application for a highly difficult new medical technology in-house, we compared the surgical outcomes (median values) of 15 patients who had undergone total hysterectomy at our hospital using the da Vinci SP (1-port) system (SP group) for uterine myoma after March 2023 and of 154 patients who underwent total hysterectomy using the conventional da Vinci Xi (four ports) system (Xi group) for uteri weighing <500 g. RESULTS: The results of the comparison of the characteristics between 15 patients in the SP group and 154 patients in the Xi group were as follows: uterus weight (g): 230 (90-500) versus 222 (55-496) (p = .35); surgical time (minutes): 199 (171-251) versus 198 (88-387) (p = .63); intraoperative blood loss (mL): 13 (5-82) versus 20 (2-384) (p = .17); and rate of surgical complication (%): 0.0 versus 1.3 (p = .66). The data indicated a comparable weight of the resected uterus, surgical time, intraoperative blood loss, and rate of surgical complications between the two groups. CONCLUSION: Robotic-assisted total hysterectomy using the da Vinci SP surgical system allowed clinicians to safely perform surgeries according to the conventional systems.

Abstract Purpose In the context of in vitro fertilization–embryo transfer (IVF–ET), factors other than egg quality may be key determinants of treatment success, in particular, maternal factors related to uterine endometrial receptivity and unidentified factors. We therefore aimed to analyze the metabolome and microbiome in IVF–ET patients who did and did not achieve pregnancy. Methods Cervicovaginal mucus was collected from patients undergoing IVF–ET. Metabolite analysis was conducted by liquid chromatography‐mass spectrometry and the microbiota were determined by the polymerase chain reaction using universal 16S‐rRNA gene bacterial primers by MiSeq sequencing. Patients were classified as pregnant (N = 10) or nonpregnant (N = 13). Metabolic pathways were examined by MetaboAnalyst. Results Three metabolic pathways, including alanine‐aspartate–glutamate metabolism, arginine biosynthesis, and cysteine‐methionine metabolism, were commonly decreased at the time of embryo transfer irrespective pregnant outcomes. Notably, pyruvate was decreased in the pregnant group. Amino acid metabolites showed inverse correlations with the presence of anaerobic microbiota in the nonpregnant group. Conclusions Metabolism decreased during embryo transplantation, with a notable decrease in pyruvate metabolism, particularly in patients who became pregnant. The behavior of metabolites in the pregnant and nonpregnant groups suggests that metabolome analysis in the cervicovaginal mucus may be a diagnostic marker for predicting pregnancy.