Down-regulation of OIP5-AS1 inhibits obesity-induced myocardial pyroptosis and miR-22/NLRP3 inflammasome axis.

BACKGROUND: Obesity can induce myocardial pyroptosis, but the exact mechanism is still unknown. A recent study reported the association of opa-interacting protein 5-antisense transcript 1 (OIP5-AS1), an evolutionarily conserved long noncoding RNA, with pyroptosis. Therefore, this study aimed to investigate the role of OIP5-AS1 in obesity-induced myocardial pyroptosis. METHODS: OIP5-AS1 was downregulated in H9c2 cells, followed by treatment with 400 µM palmitic acid (PA). Propidium iodide (PI) staining, lactic dehydrogenase (LDH) release assay, caspase-1 activity assay, IL-1ß, and IL-18 activity assay were performed to detect pyroptotic phenotype. The interaction between OIP5-AS1 and microRNAs (miRNAs) was analyzed using RNA pull-down and luciferase assay. The effect of OIP5-AS1 knockdown in high-fat diet (HFD)-induced obesity rat on cardiac function, myocardial hypertrophy, fibrosis, and remodeling was evaluated. RESULTS: Fat deposition was observed in cardiomyocytes 24 h after PA treatment; moreover, PA-treated cardiomyocytes showed significant increase in the rate of pyroptotic cells, release of LDH, protein expressions of NLRP3 and cleaved caspase-1, and the activity of caspase-1, IL-1ß, and IL-18 as well as OIP5-AS1 expression. These findings suggested that PA activated pyroptosis and induced OIP5-AS1 expression in cardiomyocytes. Moreover, OIP5-AS1 knockdown inhibited PA-induced pyroptosis. Mechanistically, OIP5-AS1 was found to specifically bind to miR-22 and to regulate NLRP3 inflammasome-mediated pyroptosis via miR-22. Furthermore, OIP5-AS1 knockdown ameliorated HFD-induced cardiac dysfunction, myocardial hypertrophy, fibrosis, remodeling, and pyroptosis. CONCLUSION: Our results revealed that downregulation of OIP5-AS1 can inhibit obesity-induced myocardial pyroptosis via miR-22/NLRP3 inflammasome axis. This finding lays a foundation of gene therapy for heart disease targeting OIP5-AS1.

Using 2D/3D ultrasound observation of endometrial thickness, endometrial volume, and blood flow changes to predict the outcome of frozen embryo transfer cycles: a prospective study.

Background: Endometrial thickness (EMT) and blood flow are commonly used indicators of endometrial receptivity. However, the results of single ultrasound examination studies differ. Therefore, we used 3-dimensional (3D) ultrasound to study the influence of changes in EMT, endometrial volume, and endometrial blood flow on frozen embryo transfer cycles. Methods: This was a prospective cross-sectional study. Women who underwent in vitro fertilization (IVF) at Dalian Women and Children's Medical Group and met the inclusion criteria were enrolled between September 2020 and July 2021. Ultrasound examinations were performed in patients who underwent frozen embryo transfer cycles on the day of progesterone administration, the third day after progesterone administration, and the day of embryo transplantation. Two-dimensional ultrasound was used to record EMT, 3D ultrasound was used to record endometrial volume, and 3D power Doppler ultrasound imaging was used to record the following endometrial blood flow parameters: vascular index, flow index, and vascular flow index. Changes in the 3 inspections of the EMT, volume, vascular index, flow index, and vascular flow index and 2 inspections of estrogen levels were categorized as "declining" or "nondeclining". The relationship between changes in a certain indicator and the IVF outcome was analyzed by univariate analysis and multifactorial stepwise logistic regression. Results: In total, 133 patients were enrolled in this study, 48 were excluded, and 85 were included in the statistical analysis. Among these 85 patients, 61 were pregnant (71%), 47 had a clinical pregnancy (55%), and 39 had an ongoing pregnancy (45%). The results showed that if the first change in endometrial volume was nondeclining, the outcomes of clinical and ongoing pregnancies were more likely to be unfavorable (P=0.03, P=0.01). Additionally, if the second change in endometrial volume on the day of embryo transplantation was nondeclining, the outcome of an ongoing pregnancy was more likely to be favorable (P=0.03). Conclusions: The changes in endometrial volume was a factor helpful in predicting the IVF outcome, whereas the changes in EMT and endometrial blood flow analyses were not helpful in predicting IVF outcome.

Quantification of cervical stiffness changes in single and twin pregnancies using the E-Cervix technique.

BACKGROUND: Cervical length is a commonly used clinical indicator for evaluating preterm birth. However, some studies have shown that cervical length does not change in patients with cervical insufficiency. OBJECTIVE: This study aimed to use the transvaginal ultrasound E-Cervix to quantitatively evaluate the cervical stiffness in women with singleton and twin pregnancies to provide a reference for the clinical prediction of preterm birth. STUDY DESIGN: We collected the cervical length, cervical hardness ratio, mean internal os strain, and mean external os strain using transvaginal ultrasonographic E-Cervix assessments in pregnant women undergoing routine examinations in the obstetrics department of our hospital from January 2020 to December 2020. We summarized the range of cervical elasticity parameters at different gestational ages and compared the cervical elasticity parameters between singleton and twin pregnancies and between preterm births and full-term births. RESULTS: A total of 988 pregnant women were enrolled in this study, and after exclusion, 770 pregnant women were enlisted; the interrater and internal consistency for various elasticity parameters were favorable; and cervical elasticity changes in women at full term showed some particular patterns. The declines in cervical length and hardness ratio were commensurate with gestational age, whereas the internal os strain and external os strain rose with increasing gestational age. The cervical hardness ratio in women with twin pregnancies was lower than in women with singleton pregnancies at the same gestational age, although the internal os strain in women with twin pregnancies was higher than in women with singleton pregnancies at the same gestational age (P<.05). The cervical length of women with twin pregnancies was shorter than that of women with singleton pregnancies at the same gestational age when the gestational age was ≥28 weeks of gestation (all P<.05), and the hardness ratio was linearly correlated with gestational age (r=0.68 and r=0.71). The regression model for the cervical hardness ratio in women with singleton pregnancies was hardness ratio = -0.8764×gestational age+100.99, whereas the regression model for the cervical hardness ratio in women with twin pregnancies was hardness ratio = -1.3037×gestational age+103.03. When we compared cervical elasticity parameters between preterm and full-term births, we noted that the cervical hardness ratio in pregnant women who exhibited preterm births was lower regardless of whether they carried singleton or twin pregnancies (P=.000 and P=.000), although their internal os strain was higher (P=.023 and P=.000). We observed no significant difference when we compared the cervical length and external os strain of pregnant women manifesting preterm births vs women with full-term births (P=.216 and P=.345 vs P=.475 and P=.363). CONCLUSION: When used for the quantification of cervical hardness, the E-Cervix cervical elasticity changes in pregnant women at full term showed some particular patterns. Cervical length and hardness ratio diminished as gestational age increased, whereas internal os strain and external os strain rose as gestational age increased; moreover, the degree of these changes was greater in women with twin pregnancies than in women with singleton pregnancies. Pregnant women experiencing preterm births exhibited a reduced cervical hardness ratio and augmented internal os strain relative to women experiencing full-term births.

A Study on the Prediction of Reproductive Outcomes in Frozen Embryo Transfer Cycles by Calculating the Volume of Uterine Junctional Zone with Three-Dimensional Ultrasound.

PURPOSE: To prospectively study the influence of the volume of the uterine junctional zone (JZ) as a novel predictor of reproductive outcomes in frozen embryo transfer cycles. METHODS: Among the first 30 patients, intra- and interobserver repeatability was evaluated and expressed as a coefficient of repeatability. The same classification system was used to evaluate the JZ of 142 infertility patients undergoing in vitro fertilization (IVF). Ultrasonography was performed on the day before transplantation. The three-dimensional (3D) volume images were then analyzed to obtain the volume of the endometrium (EV), the average thickness of the JZ on the coronal plane, and the volume of the JZ (JZV). The JZV was then divided by the EV. These parameters were compared with the outcomes of clinical pregnancy. RESULTS: The 3D image showed that the JZ achieved a good intra- and interobserver consistency (k = 0.862, k = 0.694). The total pregnancy rate was 47%. There was a highly significant difference between pregnant and non-pregnant women with respect to age (p < 0.001), JZV (p = 0.003), and JZV/EV (p < 0.001) on the day before transplantation. Age and JZV/EV were independent factors for predicting the success of IVF transplantation (p = 0.010, p = 0.016). The area under the ROC curve of JZV/EV in predicting clinical pregnancy was 0.688, the cut-off value was 0.54, the sensitivity was 83.8%, and the specificity was 50.0%. CONCLUSION: Age and JZV/EV are independent factors for predicting the success of frozen embryo transfer cycles in IVF. A smaller JZV/EV was more beneficial for clinical pregnancy.

Sensitivity of contrast-enhanced transthoracic echocardiography for the detection of residual shunts after percutaneous patent foramen ovale closure.

The objective of this study is to investigate the sensitivity of contrast-enhanced transthoracic echocardiography (c-TTE) versus transesophageal echocardiography (TEE) to detect a residual right-to-left shunt (RLS) following a transcatheter patent foramen ovale (PFO) closure.A self-controlled study was conducted in a tertiary referral hospital. 57 patients with PFO who had experienced migraines underwent percutaneous PFO closure. c-TTE, TEE, and contrast-enhanced transcranial Doppler (c-TCD) at resting and Valsalva maneuver were performed during the 3-month follow-up after the closure.The closure devices were successfully implanted in all patients without complications. Three months after closure, TEE did not detect residual Valsalva shunts in any of the 57 patients; residual valsalva shunts were found via c-TTE in 15 of the 57 patients and were also observed via c-TCD. When c-TTE and/or c-TCD were used, the rate of residual RLSs detected in patients who underwent PFO closure was 26.32%, which was significantly different than the rate detected using TEE (P < .05)c-TTE and c-TCD showed equivalent sensitivity in evaluating transcatheter closure of a PFO. c-TTE could be a more cost-effective and reliable method to detect the residual shunt after PFO closure.

Downregulation of growth arrest­specific transcript 5 alleviates palmitic acid­induced myocardial inflammatory injury through the miR­26a/HMGB1/NF­κB axis.

Palmitic acid (PA) can induce lipotoxic damage to cardiomyocytes, although its precise mechanism of action has not been completely elucidated. Growth arrest­specific transcript 5 (GAS5) is a long noncoding RNA that serves a regulatory role in several pathological processes, including tumorigenesis, stroke, cardiac fibrosis and osteoarthritis; however, its role in PA­induced myocardial injury remains elusive. The present study aimed to explore the role and underlying mechanism of GAS5 on PA­induced myocardial injury. The expression of GAS5 in PA­treated cardiomyocytes (H9c2 cells) was detected by reverse transcription­quantitative polymerase chain reaction (RT­qPCR), and its effects on PA­induced myocardial injury were measured by Cell Counting Kit­8 and lactate dehydrogenase (LDH) assays. The activities of cytokines and nuclear factor (NF)­κB were also detected by enzyme­linked immunosorbent assay, while interactions between GAS5 and microRNA (miR)­26a were evaluated by luciferase reporter assay and RT­qPCR. The regulation of GAS5 on high mobility group box 1 (HMGB1) expression was detected by RT­qPCR and western blotting. The results demonstrated that GAS5 was significantly upregulated in cardiomyocytes following treatment with PA. GAS5­knockdown increased the viability of PA­treated cardiomyocytes and reduced the activity of LDH, tumor necrosis factor­α and interleukin­1ß. Furthermore, the present study identified that GAS5 specifically binds to miR­26a, and a reciprocal negative regulation exists between the two. The present study also demonstrated that GAS5 downregulation inhibited HMGB1 expression and NF­κB activation, while these suppressive effects were mediated by miR­26a. In conclusion, the present study demonstrated that PA can induce GAS5 expression and that the downregulation of GAS5 alleviated PA­induced myocardial inflammatory injury through the miR­26a/HMGB1/NF­κB axis. These data may provide a novel insight into the mechanism of myocardial lipotoxic injury.