Transcriptome analysis identification of A-to-I RNA editing in granulosa cells associated with PCOS.

Background: Polycystic ovary syndrome (PCOS) is a complex, multifactor disorder in women of reproductive age worldwide. Although RNA editing may contribute to a variety of diseases, its role in PCOS remains unclear. Methods: A discovery RNA-Seq dataset was obtained from the NCBI Gene Expression Omnibus database of granulosa cells from women with PCOS and women without PCOS (controls). A validation RNA-Seq dataset downloaded from the European Nucleotide Archive Databank was used to validate differential editing. Transcriptome-wide investigation was conducted to analyze adenosine-to-inosine (A-to-I) RNA editing in PCOS and control samples. Results: A total of 17,395 high-confidence A-to-I RNA editing sites were identified in 3,644 genes in all GC samples. As for differential RNA editing, there were 545 differential RNA editing (DRE) sites in 259 genes with Nucleoporin 43 (NUP43), Retinoblastoma Binding Protein 4 (RBBP4), and leckstrin homology-like domain family A member 1 (PHLDA) showing the most significant three 3'-untranslated region (3'UTR) editing. Furthermore, we identified 20 DRE sites that demonstrated a significant correlation between editing levels and gene expression levels. Notably, MIR193b-365a Host Gene (MIR193BHG) and Hook Microtubule Tethering Protein 3 (HOOK3) exhibited significant differential expression between PCOS and controls. Functional enrichment analysis showed that these 259 differentially edited genes were mainly related to apoptosis and necroptosis pathways. RNA binding protein (RBP) analysis revealed that RNA Binding Motif Protein 45 (RBM45) was predicted as the most frequent RBP binding with RNA editing sites. Additionally, we observed a correlation between editing levels of differential editing sites and the expression level of the RNA editing enzyme Adenosine Deaminase RNA Specific B1 (ADARB1). Moreover, the existence of 55 common differentially edited genes and nine differential editing sites were confirmed in the validation dataset. Conclusion: Our current study highlighted the potential role of RNA editing in the pathophysiology of PCOS as an epigenetic process. These findings could provide valuable insights into the development of more targeted and effective treatment options for PCOS.

Epitranscriptomic investigation of myopia-associated RNA editing in the retina.

Myopia is one of the most common causes of vision loss globally and is significantly affected by epigenetics. Adenosine-to-inosine (A-to-I RNA) editing is an epigenetic process involved in neurological disorders, yet its role in myopia remains undetermined. We performed a transcriptome-wide analysis of A-to-I RNA editing in the retina of form-deprivation myopia mice. Our study identified 91 A-to-I RNA editing sites in 84 genes associated with myopia. Notably, at least 27 (32.1%) of these genes with myopia-associated RNA editing showed existing evidence to be associated with myopia or related ocular phenotypes in humans or animal models, such as very low-density lipoprotein receptor (Vldlr) in retinal neovascularization and hypoxia-induced factor 1 alpha (Hif1a). Moreover, functional enrichment showed that RNA editing enriched in FDM was primarily involved in response to fungicides, a potentially druggable process for myopia prevention, and epigenetic regulation. In contrast, RNA editing enriched in controls was mostly involved in post-embryonic eye morphogenesis. Our results demonstrate altered A-to-I RNA editing associated with myopia in an experimental mouse model and warrant further study on its role in myopia development.

Epidemiological features and management of eye burn patients in Wuxi, China.

BACKGROUND: The current study aimed to analyse epidemiological data on eye burns in Wuxi, China, for the years 2015-2021, and to provide insight into the development of appropriate prevention strategies. METHODS: A retrospective study was conducted on 151 hospitalised patients with eye burns. Data collected included gender, age, the monthly distribution of incidence, cause of eye burn, the site of eye burn, the type of surgery, visual outcome, the length of hospital stay and the cost of hospital admission. Statistical analysis was performed using SPSS V.19.0 and Graph Pad Prism V.9.0. RESULTS: In a total of 151 eye burn patients, 130 were males (86.09%) and 21 were females (13.91%). The proportion of patients classified as grade III was the greatest (46.36%). The average age of our hospitalised patients with eye burns was 43.72 years and the average length of hospital stay was 17 days. The number of injuries was highest in September (14.6%). Among eye burn patients, workers and farmers became the most common occupations (62.91%, 12.58%). The most frequent cause of burns was alkali burns (19.21%), followed by acid burns (16.56%). When admitted to the hospital, patients' average vision was 0.06, and 49% of them had a poor vision (<0.3, ≥0.05). CONCLUSION: With an investigation of 7-year hospitalisation data, the current study provided a fundamental reference for epidemiological features and management of eye burns in Wuxi, China, which could contribute to the development of treatment and prevention strategies.

Quantitative Analysis on Cartilage Growth Between Ipsilateral and Contralateral Donor Sites in Microtia Patients.

BACKGROUND: Costal cartilage harvest is required in patients with unilateral microtia when autologous reconstruction is being considered. However, whether an ipsilateral or contralateral donor site should be used remains controversial. This is the first study to compare cartilaginous growth between ipsilateral and contralateral donor sites in patients with unilateral microtia. METHODS: In this retrospective study of 58 patients, the lengths of the sixth to ninth costal cartilages and 3 position-defining measurements with respect to the sixth to ninth costochondral junctions were calculated using 3-dimensional costal cartilage imaging. Patients were divided into subgroups, and the lateral differences between isolated microtia and hemifacial microsomia and between the growing and adult age groups, were compared. RESULTS: In the isolated group, the sixth and seventh costal cartilages were longer on the contralateral side. The transverse dimension on the contralateral side, with respect to the sixth and seventh costochondral junctions, was also larger than that on the ipsilateral side in growing patients. However, no significant difference was observed between the 2 sides in the hemifacial microsomia group; there was also no difference between the age-related groups in this regard (P > 0.05). CONCLUSIONS: These findings suggest that age- and side-related differences in donor sites should be considered in patients with isolated microtia.

Decreased RNA m6A methylation enhances the process of the epithelial mesenchymal transition and vasculogenic mimicry in glioblastoma.

RNA N6-methyladenosine (m6A) modification is gradually thought to be an active participant in the considerable biological processes of glioblastoma (GB), providing us with a novel insight for exploring this disease. However, the role of RNA m6A modification during the epithelial mesenchymal transition (EMT) or vasculogenic mimicry (VM) progression has not been investigated in GB. Here we performed a research to validate the impact exerted by AlkB homolog 5 (ALKBH5), one of "erasers" for RNA m6A and methyltransferase-like 3 (METTL3) which adds m6A modification to the RNAs on the progression of EMT and VM in GB. In this study, we demonstrate that the m6A levels of RNAs were reduced in GB cells and glioma tissues. Patients with high mRNA expression of ALKBH5 acquired relatively shorter median overall survival (OS) time, while patients with relatively high expression of MEETL3 prolonged their disease free survival. ALKBH5 enhanced GB cell proliferation and influenced cell cycle in vitro. Decreased RNA m6A methylation enhanced the progression of the EMT and VM in glioblastoma cells. ALKBH5 strengthened glioblastoma growth and enhanced the EMT and VM process of glioblastoma in vivo. Our study uncovers that RNA m6A methylation suppresses the process of EMT and VM in glioblastoma, providing a new perspective to seek for a potential therapeutic target for GB.

One-step simultaneous quantitative detection of three pesticides based on bimetallic organic framework nanomaterials and aptamers.

We have developed a new method of one-step simultaneous detection for three pesticides including acetamiprid, atrazine and carbendazim based on organic framework nanomaterial Cu/UiO-66 and three different fluorescent dyes labeled pesticide aptamers. Cu/UiO-66 can be easily combined with pesticide aptamers through strong coordination, and then aptamers were adsorbed to the surface of Cu/UiO-66, which brings the dyes and Cu/UiO-66 into close proximity. Then, the fluorescence of dyes was quenched by Cu/UiO-66. When the target pesticides appeared, the aptamers reacted with corresponding target pesticides and formed special spatial structure, and then the dyes were far away from the surface of Cu/UiO-66 and the fluorescence of dyes is resumed. Thus, the one-step simultaneous detection for three pesticides can be achieved by synchronous fluorescence analysis. The detection limit of acetamiprid, atrazine and carbendazim were 0.1 nmol/L, 1.6 nmol/L, and 0.3 nmol/L, respectively. This method has a good sensitivity, low detection limit, and high selectivity. We have developed a new method of one-step simultaneous detection for three pesticides including acetamiprid, atrazine and carbendazim based on a multi-color fluorescent probe composed of bimetallic organic framework nanomaterials Cu/UiO-66 and three different fluorescent dyes and phosphate double-labeled aptamers of acetamiprid, atrazine and carbendazim.

Validity and reliability of three-dimensional costal cartilage imaging for donor-site assessment and clinical application in microtia reconstruction patients: A prospective study of 22 cases.

OBJECTIVES: This study assesses the ability to reconstruct costal cartilage images by using three-dimensional visualisation software (Mimics) based on semi-automated segmentation algorithm and to investigate its reliability and validity with an anthropometric analysis. DESIGN: Observational prospective study. SETTING: Plastic surgery department of a tertiary hospital. PARTICIPANTS: Twenty-two microtia patients who underwent autologous ear reconstruction. MAIN OUTCOME MEASURES: Preoperative thoracic computed tomography data were processed to Mimics software for three-dimensional costal cartilage imaging. The length, width, thickness and volume of the 9th costal cartilages were calculated from these images and compared with the direct measurements (DM) obtained intraoperatively. RESULTS: The intra-examiner reliability and inter-examiner reliability were high in terms of all four measurements (intraclass correlation coefficients, ICC: 0.876-0.984). There were no significant differences between image-based anthropometry and DM in the linear measurements except for the volume (P < .05). The mean volume calculation error of Mimics was -0.08 ± 0.13 mL. No correlation was found between the anthropometric variables and the absolute errors (P > .05). Furthermore, Bland-Altman plots were used to evaluate the agreement between the two methods. CONCLUSIONS: Despite a very small error was found in volume calculation, Mimics software was accurate and reliable in linear calculation. Three-dimensional costal cartilage imaging was found to be an efficient tool for morphological evaluation of costal cartilages. We believe that with the application of individualised cartilage models based on three-dimensional printing, the use of customised ear framework carving will be practicable in surgical training.

[Effect of adipose tissue extract on promoting angiogenesis and adipogenesis in tissue engineering chamber in vivo].

OBJECTIVE: To evaluate the influence of adipose tissue extract on inducing angiogenesis and adipogenesis in adipose tissue engineering chamber in vivo. METHODS: 6 months' healthy New Zealand rabbits (n = 64) were picked. The inguinal fat pads were cultured, centrifuged, filtered, and the liquid was called adipose tissue extract (ATE). Two adipose tissue engineering chamber were built in the rabbit's back. A week later, 0.2 ml normal saline (control group, left) and 0. 2 ml ATE (experimental group, right) was respectively injected into the chamber. The contents were evaluated morphometrically, histologically and immunohistochemically 3 days, 1 week, 2 weeks, 3 weeks, 4 weeks and 7 weeks after injection. 8 rabbits were observed each time. The data regarding the number of the volume of fat flap and blood capillary at each time point were analyzed by paired t test. RESULTS: After injection, new tissue volume was significantly increased in the experimental group [(5.12 ± 0.22) ml], compared with that in control group [(4.90 ± 0.15) ml]. Early angiogenesis was also increased after ATE injection and the total number of capillaries reached peak 1 week after injection, which was (72.80 ± 9.67) in experimental group and (51.40 ± 6.09) in control group. In the mid-term of experimental period, earlier adipogenesis appeared in experimental group. In the later period, the outer capsule of the new construction was thinner in experimental group which reduced the suppression of the adipogenesis. CONCLUSIONS: ATE can promote the angiogenesis and adipogenesis in the chamber, and reduce the capsule contracturing, so as to induce the large volume of adipose tissue regeneration

Role of oxidative stress in the promoting activities of pcbs.

PCBs are organic pollutants that persist and bioaccumulate in the environment. These chemicals induce and promote liver tumors in rodents. Previous studies have shown that they increase oxidative stress in the liver, including lipid peroxidation, oxidative DNA damage, and NF-κB activation. The objective of these studies was to determine if the promoting activities of PCBs could be inhibited by dietary antioxidants (vitamin E, selenium, or phytochemicals) or by knocking out the p50 subunit of NF-κB. In the antioxidant studies, female rats were first injected with DEN (150 mg/kg) and then administered 4 biweekly i.p. injections (300 µmol/kg/injection) of PCB-77, PCB-153, or vehicle; the number and volume of placental glutathione S-transferase (PGST)-positive foci were then quantified. Vitamin E did not influence the promoting activities of PCBs. Increasing dietary selenium above the recommended intake increased the number of foci induced but decreased their volume. Most of the phytochemicals examined (N-acetyl cysteine, ß-carotene, resveratrol, EGCG) had no significant effect on the promoting activity of PCB-77. Ellagic acid increased and lycopene decreased the number of foci; ellagic acid, CoQ(10), and curcumin decreased the volume of foci. In the NF-κB knockout study, male mice were first injected with DEN (90 mg/kg); controls not receiving DEN were also studied. Both p50 -/- and wild-type mice were then injected biweekly 20 times with PCB-153 (300 (µmol/kg). In DEN-treated and DEN + PCB-treated mice, the incidence of tumors was lower in the p50 -/- mice than in wild-type mice. In mice receiving PCB-153, the tumor incidence and tumor volume were higher. The volume of tumors that were positive for glutamine synthetase was increased in mice administered PCB-153. This study shows that the promotion of hepatocarcinogenesis by PCBs is largely unaffected by dietary antioxidants but is diminished when NF-κB activation is impaired by the absence of the p50 subunit.

Dietary vitamin E does not inhibit the promotion of liver carcinogenesis by polychlorinated biphenyls in rats.

In this study, the effect of dietary vitamin E on the hepatic tumor-promoting activity of PCB-77 and PCB-153 in female Sprague-Dawley rats (175-200 g) was investigated. One week after diethylnitrosamine injection, rats were fed purified diets containing 10, 50, or 250 mg/kg vitamin E in the form of alpha-tocopheryl acetate. Starting 1 wk later, we injected rats i.p. with vehicle (corn oil) or PCB-77 or PCB-153 (300 mumol/kg) every 14 d for 4 injections. All rats were killed 10 d after the last PCB injection. The number and volume of placental glutathione S-transferase (PGST)-positive foci were increased by PCB-77 but not by PCB-153. Vitamin E did not affect the induction of PGST-positive foci. PCB-77, but not PCB-153, increased hepatic NF-kappaB activity. In conclusion, dietary vitamin E supplementation does not protect against the induction of altered hepatic focal lesions by PCBs.

Effect of 2,2',4,4',5,5'-hexachlorobiphenyl (PCB-153) on hepatocyte proliferation and apoptosis in mice deficient in the p50 subunit of the transcription factor NF-kappaB.

Polychlorinated biphenyls (PCBs) are a group of synthetic chemicals that induce and promote liver tumors in rodents. We previously showed hepatic nuclear factor kappaB (NF-kappaB) activation and increased hepatocyte proliferation in PCB-treated rats. In this study, the role of NF-kappaB in hepatocyte proliferation and apoptosis after PCB administration was analyzed in wild-type mice and in mice deficient in the NF-kappaB p50 subunit (p50-/-). In a 2-day study, mice received a single intraperitoneal (ip) injection of corn oil or PCB-153. Hepatic NF-kappaB DNA binding activity and cell proliferation were increased by PCB-153 in wild-type mice but not in p50-/- mice. In a 21-day study, mice received six ip injections of corn oil or PCB-153 (twice weekly for 3 weeks) and were euthanized 4 days after the last injection. In this study, NF-kappaB DNA binding activity was not increased after PCB-153 treatment in wild-type or p50-/- mice. Cell proliferation was significantly increased in the wild-type mice treated with PCB-153; in the p50-/- mice treated with PCB-153, cell proliferation was greater than in untreated mice but less than in wild-type mice treated with PCB-153. The livers of p50-/- mice showed greater apoptosis than those of wild-type mice; PCB-153 decreased apoptosis in p50-/- mice, with higher inhibition in the 21-day study than in the 2-day study. RNase protection assays indicated that PCB-153 decreased the mRNA level of cyclin A2, B1, B2, and C in the 2-day study, but not in the 21-day study; however, it did not affect cyclin D1 and D2 mRNA levels at either time point. Cyclin D1 protein levels were not affected by PCB-153. Taken together, these data indicate that the absence of the NF-kappaB p50 subunit alters the proliferative and apoptotic changes in mouse liver in the response to PCB-153.

Effect of a single dose of polychlorinated biphenyls on hepatic cell proliferation and the DNA binding activity of NF-kappaB and AP-1 in rats.

Polychlorinated biphenyls (PCBs) are environmental pollutants that, because of their persistence and biomagnification, raise concerns about the health consequences of long-term exposure. PCB mixtures induce hepatocellular carcinomas in rodents, but the mechanism of their promoting activity is not clear. Previous studies have shown that oxidative stress occurs after PCB administration, with the induction of lipid peroxidation and oxidative DNA damage, which may contribute to their promoting activity. In this study, we examined whether the oxidative stress-sensitive transcription factors NF-kappaB or AP-1 were activated by PCBs in the liver. Male Sprague-Dawley rats were injected i.p. with corn oil, 2,2',4,4',5,5'-hexachlorobiphenyl (PCB-153, 30, 150, or 300 micromol/kg), 3,3',4,4'-tetrachlorobiphenyl (PCB-77, 30, 150, or 300 micromol/kg), or both PCBs (each 30 or 150 micromol/kg). Rats were euthanized 2, 6, or 24 h, or 2, 6, and 10 d after the PCB injection. Electrophoretic mobility shift assays (EMSAs) were performed to determine NF-kappaB and AP-1 DNA binding activities. The highest NF-kappaB DNA binding activity was observed in rats receiving higher doses of PCB-153 (150 and 300 micromol/kg), with peak activation occurring 2 d after injection. AP-1 activation was not detected at any timepoint. Hepatocyte proliferation, as measured by the labeling index, was increased only in groups receiving the highest dose of PCB-153 or the combination of two PCBs (150 micromol/kg each) at day 2, and not by any other PCB treatment at any timepoint. These results show that PCB-153, but not PCB-77, can induce hepatocyte proliferation and hepatic NF-kappaB activation after a single dose.