Cellular Functions of High-Temperature Requirement Factor A4 in Placenta.

The expression of High-temperature requirement factor A4 (HtrA4) mRNA is significantly lower in the chorionic villi of patients with recurrent pregnancy loss (RPL) than in the control group. We conducted an investigation into the cellular functions of HtrA4 using the CRISPR/Cas9 system and shRNA-HtrA4 to create knockout BeWo cells and HtrA4 knockdown JEG3 cells. Our results indicated that the knockout BeWo cells exhibited reduced capacity for invasion and fusion, but increased levels of proliferation and migration, with a significantly shortened cell cycle compared to wild-type cells. Wild-type BeWo cells highly expressed cell invasion- and fusion-related factors, while knockout BeWo cells highly expressed migration-, proliferation-, and cell cycle-related factors. The shRNA-HtrA4 JEG3 cells showed a decreased capacity for invasion, but an increased capacity for migration, accompanied by a decrease in the expression of cell invasion-related factors and an increase in migration-related factors. Moreover, our ELISA results revealed that the serum HtrA4 level was lower in patients with RPL than in the controls. These findings suggest that HtrA4 depletion may be associated with placental dysfunction.

Hepatitis B surface antigen and antibody positivity among women of childbearing age after three decades of universal vaccination in South Korea.

OBJECTIVES: We estimated the impact of universal hepatitis B immunization using 18-year data of women who are of childbearing age in South Korea. METHODS: We used hepatitis B surface antigen (HBsAg) and antibody (anti-HBs) data of 145,993 women aged 20-49 years during 2001-2018 at the Gangnam CHA Medical Center. Annual prevalences of HBsAg and anti-HBs positivity were calculated and tested for linear trend. We conducted age-period-cohort (APC) analysis to obtain period and cohort effect. RESULTS: Overall proportion of HBsAg positivity was 3.5% (n = 5050) and anti-HBs positivity was 75.3% (n = 109,907) during the study period. HBsAg positivity percentage decreased from 5.1% in 2001 to 2.5% in 2018 (P < 0.001) while anti-HBs positivity increased from 59.9% to 75.8% (P = 0.002). Average annual percent change of HBsAg positivity was -5.9% (95% confidence interval (CI): -6.9%, -4.8%). The period and cohort RR curve identified a consistent decrease in HBsAg positivity over time and across generations. CONCLUSIONS: We observed a concurrent decrease in HBsAg and an increase in anti-HBs seropositivity among Korean women of childbearing age, implicating success in preventing vertical transmission.

Peripheral blood natural killer cell proportion and ovarian function in women with recurrent implantation failure.

Current knowledge of the association between peripheral natural killer (NK) cell proportion and ovarian function in reproductive-age women is limited. We explored the association between NK cell proportion and ovarian function in women who underwent in-vitro fertilization (IVF) treatment. This was a retrospective cohort study using the data of 20-44-year-old women with recurrent implantation failure (RIF) who were tested for NK cell proportion and anti-Müllerian hormone (AMH). Indicators of ovarian function included AMH, observed-to-(age-appropriate) reference AMH ratio, high FSH, peak E2 and total number of oocytes during the first IVF cycle following the test. We used different model specification controlling for women's age, and body mass index. Among a total of 936 women, majority showed lower AMH compared to age-appropriate level. Average NK cell proportion was 13.5 ± 5.7%. Number of oocytes showed positive association with NK cell (ß = 0.040, p = .025). In the subgroup with NK ≥ 18%, NK cell proportion was negatively associated with AMH (-0.106, p = .012), AMH ratio (-0.049, p = .014) and number of oocytes (-0.021, p < .001) while the associations with others remain close to null. High NK cell proportion may be harmful to ovarian reserve or function.

3D-cultured human placenta-derived mesenchymal stem cell spheroids enhance ovary function by inducing folliculogenesis.

Placenta-derived mesenchymal stem cells (PD-MSCs) have numerous advantages over other adult MSCs that make them an attractive cell source for regenerative medicine. Here, we demonstrate the therapeutic effect of PD-MSCs in ovariectomized (Ovx) rats and compare their efficacy when generated via a conventional monolayer culture system (2D, naïve) and a spheroid culture system (3D, spheroid). PD-MSC transplantation significantly increased the estradiol level in Ovx rats compared with the non-transplantation (NTx) group. In particular, the estradiol level in the Spheroid group was significantly higher than that in the Naïve group at 2 weeks. Spheroid PD-MSCs exhibited a significantly higher efficiency of engraftment onto ovarian tissues at 2 weeks. The mRNA and protein expression levels of Nanos3, Nobox, and Lhx8 were also significantly increased in the Spheroid group compared with those in the NTx group at 1 and 2 weeks. These results suggest that PD-MSC transplantation can restore ovarian function in Ovx rats by increasing estrogen production and enhancing folliculogenesis-related gene expression levels and further indicate that spheroid-cultured PD-MSCs have enhanced therapeutic potential via increased engraftment efficiency. These findings improve our understanding of stem-cell-based therapies for reproductive systems and may suggest new avenues for developing efficient therapies using 3D cultivation systems.

Opposing roles of inter-α-trypsin inhibitor heavy chain 4 in recurrent pregnancy loss.

BACKGROUND: The mechanism behind an increased risk of recurrent pregnancy loss (RPL) remains largely unknown. In our previous study, we identified that inter-α-trypsin inhibitor heavy chain 4 (ITI-H4) is highly expressed at a modified molecular weight of 36 kDa in serum derived from RPL patients. Yet, the precise molecular mechanism and pathways by which the short form of ITI-H4 carries out its function remain obscure. METHODS: Human sera and peripheral blood mononucleated cells (PBMCs) were collected from patients and normal controls to compare the expression levels of ITI-H4 and plasma kallikrein (KLKB1). Flow cytometric assay was performed to measure inflammatory markers in sera and culture supernatants. Furthermore, to investigate the functions of the two isoforms of ITI-H4, we performed migration, invasion, and proliferation assays. FINDINGS: In the current study, we showed that ITI-H4 as a biomarker of RPL could be regulated by KLKB1 through the IL-6 signaling cascade, indicating a novel regulatory system for inflammation in RPL. In addition, our study indicates that the two isoforms of ITI-H4 possess opposing functions on immune response, trophoblast invasion, and monocytes migration or proliferation. INTERPRETATION: The ITI-H4 (∆N688) might be a crucial inflammatory factor which contributes to the pathogenesis of RPL. Moreover, it is expected that this study would give some insights into potential functional mechanisms underlying RPL. FUND: This study was supported by the Ministry of Health & Welfare of the Republic of Korea (HI18C0378) through the Korea Health Industry Development Institute.

[The measurement of gastric emptying time with 3-D ultrasonography].

BACKGROUND/AIMS: Three dimensional (3-D) ultrasonography is definitely more accurate than conventional 2-D ultrasonography in volume measurement of intra-abdominal organs. However, its application in measuring gastric emptying time has been limited. Thus, we tried to measure gastric antral emptying time by using 3-D ultrasonography and compare with that by radio-scintigraphy. METHODS: We performed both 3-D ultrasonography and radio-scintigraphy on the same day in 23 healthy subjects. After overnight fast, the subjects ingested 500 mL of soup meal (84 Kcal) that had been pre-mixed with 1 mCi of technetium(99m). The half emptying time (T(1/2)) measured by 3-D ultrasonography was defined by the time when the gastric antral volume decreased to half. RESULTS: The mean T(1/2) of 23 healthy subjects measured by 3-D ultrasonography was 23.4 +/- 10.5 min, while that measured by scintigraphy was 28.4 +/- 14.4 min. The difference (p=0.11 by paired t-test) between the two methods was not significant. Moreover, no significant correlation of T1/2 between the two methods (r=0.361, p=0.09) was observed. CONCLUSIONS: Measuring gastric antral emptying time with 3-D ultrasonography may be useful but require further validation studies and advance in technology.