Circ-myh8 Promotes Pulmonary Hypertension by Recruiting KAT7 to Govern Hypoxia-Inducible Factor-1α Expression.

Background Aberrant expression of circular RNAs (circRNAs) contributes to the initiation and progression of pulmonary hypertension (PH). Hypoxia-inducible factor (HIF) is a well-known modulator of hypoxia-induced PH. The role and underlying mechanism of circRNAs in the regulation of HIF expression remains elusive. Methods and Results We profiled pulmonary artery transcriptomes using RNA sequencing and screened circRNAs associated with hypoxia treatment. The expression of a novel circRNA, circ_chr11_67292179-67294612 (circ-myh8), was increased by hypoxia in a time-dependent manner. We evaluated the effects of circ-myh8 overexpression by adeno-associated virus or inhibition by short hairpin RNA on proliferation and cell cycling in mice and pulmonary artery smooth muscle cells. Overexpression of circ-myh8 promotes PH under normoxia, and disruption of circ-myh8 by short hairpin RNA mitigates PH in chronic hypoxic mice. Biologically, circ-myh8 induces the proliferation and cell-cycle progression of pulmonary artery smooth muscle cells in vivo and in vitro. Mechanistically, RNA pull-down and RNA immunoprecipitation assays were used to examine the interaction of circRNAs with the binding protein KAT7 (lysine acetyltransferase 7). The acetylation level of lysine 5 of histone H4 in the transcriptional initiation region of HIF1α was determined by chromatin immunoprecipitation assay followed by reverse transcription-quantitative polymerase chain reaction. Circ-myh8 acts as a modular scaffold to recruit histone acetyltransferase KAT7 to the promoters of HIF1α, which elicits acetylation of lysine 5 of histone H4 in their promoters. Conclusions Our findings not only reveal the pivotal roles of circ-myh8 in governing histone modification in anti-PH treatment but also advocate triggering the circ-myh8/KAT7/HIF1α pathway to combat PH.

The feasibility of laparoscopically assisted, hysteroscopic removal of interstitial pregnancies: A case series.

OBJECTIVE: The study objective was to assess the feasibility of the management of interstitial pregnancy by laparoscopically assisted hysteroscopic removal. METHODS: This retrospective study included a case series of 17 patients who were diagnosed interstitial pregnancy with dilated proximal tubal ostium by transvaginal ultrasonography at the Women's hospital, School of Medicine, Zhejiang University between August 2017 and October 2020. Laparoscopically assisted hysteroscopic removals of the products of conception were performed. Various data were collected including age, surgical and obstetric history, gestational age, preoperative symptoms, human chorionic gonadotropin level and ultrasonography results. The outcomes measured were intraoperative bleeding, pathologic findings, conversions. RESULTS: Eleven cases were successfully resected the interstitial gestational products with laparoscopically assisted hysteroscopy. There were four cases failed of hysteroscopic removal, for the proximal tubal ostia were too small for the surgical instruments to enter. Then cornual wedge resections were performed. Two cases were identified as intramural pregnancy by hysteroscopic and laparoscopic view. Most of the intramural pregnancy tissue of one patient was removed by hysteroscopy. The other one converted to laparoscopy. CONCLUSION: Laparoscopically assisted hysteroscopic management could be a feasible surgical option to interstitial pregnancies. Further clinical studies are needed to establish detailed criteria to select the appropriate cases for hysteroscopic management.

Expression of flotillin-2 in human non-small cell lung cancer and its correlation with tumor progression and patient survival.

INTRODUCTION: Recent studies have revealed that flotillin-2 (FLOT2) played important roles in cancer progression. The aim of this study was to investigate the clinicopathologic and prognostic significance of FLOT2 expression in human non-small cell lung cancer (NSCLC). METHODS: Quantitative real-time PCR (qRT-PCR) was performed to detect FLOT2 mRNA expression in lung cancer cell lines, normal bronchial epithelial cells, 24 pairs of NSCLC tissues and matched adjacent non-tumor tissues. Immunohistochemistry (IHC) was performed to examine FLOT2 protein expression in paraffin-embedded tissues from 90 NSCLC patients. Statistical analyses were performed to evaluate the clinicopathological significance of FLOT2 expression. RESULTS: FLOT2 mRNA expression was evidently up-regulated in lung cancer cell lines and NSCLC tissues compared with normal bronchial epithelial cells and adjacent non-tumor tissues. In the 90 cases of tested NSCLC samples, FLOT2 protein level was positively correlated with tumor stage, and lymph node metastasis. Patients with high FLOT2 expression had shorter overall survival compared with the low FLOT2 expression group. Univariate and multivariate analyses indicated that high FLOT2 expression was an independent poor prognostic factor for NSCLC patients. CONCLUSIONS: Our findings provided that high FLOT2 expression was associated with poor outcomes in NSCLC patients, and FLOT2 could be a potential prognostic biomarker for lung cancer progression.

Expression of miR-32 in human non-small cell lung cancer and its correlation with tumor progression and patient survival.

INTRODUCTION: miR-32 has recently been found to be implicated in many critical processes in various types of human cancer. However, its clinical significance in human non-small cell lung cancer (NSCLC) has not yet been elucidated. In the present study, we investigated the expression of miR-32 in NSCLC and analyzed its association with clinical features and prognosis of NSCLC patients. METHODS: Quantitative real-time PCR (qRT-PCR) was used to measure expression level of miR-32 in lung cancer cell lines, normal bronchial epithelial cells, 90 pairs of tumor samples and adjacent non-tumor tissues. To determine its prognostic value, overall survival was evaluated using the Kaplan-Meier method. Univariate and multivariate analysis were performed using the Cox proportional hazard analysis. RESULTS: The expression of miR-32 was significantly decreased in lung cancer cell lines and NSCLC tissues compared with normal bronchial epithelial cells and adjacent non-tumor tissues (P < 0.05). This reduction of miR-32 was associated with tumor stage and lymph node metastasis (P < 0.05). Moreover, Kaplan-Meier analysis demonstrated that patients with low miR-32 expression had shorter overall survival time than those with high miR-32 expression (P < 0.05). Univariate analysis revealed statistically significant correlations between overall survival and miR-32 level, tumor stage and lymph node metastasis (P < 0.05). Furthermore, miR-32 levels, tumor stage and lymph node metastasis were independently associated with overall survival (P < 0.05). CONCLUSIONS: Our results provided the first evidence that down-regulation of miR-32 was correlated with NSCLC progression, and miR-32 might be a potential molecular biomarker for predicting the prognosis of patients.

Associations between polymorphisms in IL-12A, IL-12B, IL-12Rß1, IL-27 gene and serum levels of IL-12p40, IL-27p28 with esophageal cancer.

PURPOSE: The aim of this study was to investigate whether IL-12A, IL-12B, IL-12Rß1, and IL-27 gene polymorphisms and serum levels of IL-12, IL-27 are associated with esophageal cancer. METHODS: We genotyped IL-12A gene rs568408, IL-12B gene rs3212227, IL-12Rß1 gene 378 C/G, IL-27 gene rs153109, rs17855750, and rs181206 polymorphisms in a case-control study of 426 esophageal cancer patients and 432 health controls, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and serum IL-12p40 and IL-27p28 levels were measured by enzyme-linked immunosorbent assay. RESULTS: Both serum IL-12p40 and IL-27p28 levels were significantly higher in controls than those in patients (P < 0.01). Rs568408 AG/AA, rs3212227 CC/AC, and IL-12Rß1 378 GG/GC genotypes were associated with significantly increased risk of esophageal cancer (rs568408: χ(2) = 5.704, P = 0.017; rs3212227: χ(2) = 7.689, P = 0.006; IL-12Rß1 378C/G: χ(2) = 5.206, P = 0.023). Moreover, rs3212227 CC/AC and 378 GG/GC genotypes were observed significantly associated with decreased serum IL-12p40 level in patients compare to other genotypes (rs3212227: t = 2.129, P = 0.034; IL-12Rß1 378 C/G: t = 2.178, P = 0.030). Furthermore, frequency of rs3212227 CC/AC genotypes was significantly higher in patients with poor differentiation than those with AA genotype (χ(2) = 4.314, P = 0.035). CONCLUSION: Our data suggest that the impaired production of IL-12p40 and IL-27p28 behaves as risk factors for esophageal cancer occurrence. IL-12B gene rs3212227 CC/AC and IL-12Rß1 gene 378 GG/GC genotypes, which associated with decreased IL-12p40 level, may contribute to esophageal cancer susceptibility.