Integrated transcriptomics and metabolomics analysis provides insights into aromatic volatiles formation in Cinnamomum cassia bark at different harvesting times.

BACKGROUND: Cinnamomum cassia Presl, classified in the Lauraceae family, is widely used as a spice, but also in medicine, cosmetics, and food. Aroma is an important factor affecting the medicinal and flavoring properties of C. cassia, and is mainly determined by volatile organic compounds (VOCs); however, little is known about the composition of aromatic VOCs in C. cassia and their potential molecular regulatory mechanisms. Here, integrated transcriptomic and volatile metabolomic analyses were employed to provide insights into the formation regularity of aromatic VOCs in C. cassia bark at five different harvesting times. RESULTS: The bark thickness and volatile oil content were significantly increased along with the development of the bark. A total of 724 differentially accumulated volatiles (DAVs) were identified in the bark samples, most of which were terpenoids. Venn analysis of the top 100 VOCs in each period showed that twenty-eight aromatic VOCs were significantly accumulated in different harvesting times. The most abundant VOC, cinnamaldehyde, peaked at 120 months after planting (MAP) and dominated the aroma qualities. Five terpenoids, α-copaene, ß-bourbonene, α-cubebene, α-funebrene, and δ-cadinene, that peaked at 240 MAP could also be important in creating C. cassia's characteristic aroma. A list of 43,412 differentially expressed genes (DEGs) involved in the biosynthetic pathways of aromatic VOCs were identified, including phenylpropanoids, mevalonic acid (MVA) and methylerythritol phosphate (MEP). A gene-metabolite regulatory network for terpenoid and phenylpropanoid metabolism was constructed to show the key candidate structural genes and transcription factors involved in the biosynthesis of terpenoids and phenylpropanoids. CONCLUSIONS: The results of our research revealed the composition and changes of aromatic VOCs in C. cassia bark at different harvesting stages, differentiated the characteristic aroma components of cinnamon, and illuminated the molecular mechanism of aroma formation. These foundational results will provide technical guidance for the quality breeding of C. cassia.

Non-invasive imaging of pathological scars using a portable handheld two-photon microscope.

BACKGROUND: Pathological scars are a disorder that can lead to various cosmetic, psychological, and functional problems, and no effective assessment methods are currently available. Assessment and treatment of pathological scars are based on cutaneous manifestations. A two-photon microscope (TPM) with the potential for real-time non-invasive assessment may help determine the under-surface pathophysiological conditions in vivo . This study used a portable handheld TPM to image epidermal cells and dermal collagen structures in pathological scars and normal skin in vivo to evaluate the effectiveness of treatment in scar patients. METHODS: Fifteen patients with pathological scars and three healthy controls were recruited. Imaging was performed using a portable handheld TPM. Five indexes were extracted from two dimensional (2D) and three dimensional (3D) perspectives, including collagen depth, dermo-epidermal junction (DEJ) contour ratio, thickness, orientation, and occupation (proportion of collagen fibers in the field of view) of collagen. Two depth-dependent indexes were computed through the 3D second harmonic generation image and three morphology-related indexes from the 2D images. We assessed index differences between scar and normal skin and changes before and after treatment. RESULTS: Pathological scars and normal skin differed markedly regarding the epidermal morphological structure and the spectral characteristics of collagen fibers. Five indexes were employed to distinguish between normal skin and scar tissue. Statistically significant differences were found in average depth ( t = 9.917, P <0.001), thickness ( t = 4.037, P <0.001), occupation ( t = 2.169, P <0.050), orientation of collagen ( t = 3.669, P <0.001), and the DEJ contour ratio ( t = 5.105, P <0.001). CONCLUSIONS: Use of portable handheld TPM can distinguish collagen from skin tissues; thus, it is more suitable for scar imaging than reflectance confocal microscopy. Thus, a TPM may be an auxiliary tool for scar treatment selection and assessing treatment efficacy.

Parental chest computerized tomography examination before IVF/ICSI has no impact on pregnancy and neonatal outcomes: a cohort study of 2680 fresh transfer cycles.

BACKGROUND: Some concern has been expressed regarding the negative effects of low-level ionizing radiation exposure in the context of radiological evaluation prior to IVF/ICSI treatment, but the available evidence is limited and conflicting. The aim of this study is to evaluate pregnancy and neonatal outcomes of couples who did chest computed tomography (CT) prior to IVF/ICSI. METHODS: This was a retrospective cohort study of 2680 IVF/ICSI fresh embryo transfer cycles conducted from January 2019 - August 2020. Fertility outcomes were compared between couples that had or had not undergone CT examination within 3 months prior to the date of oocyte retrieval and sperm collection. Miscarriage was the primary study outcome, while secondary outcomes included the number of oocytes collected, oocyte maturation, normal fertilization, number of good quality cleavage stage embryos, blastocyst formation, implantation, clinical pregnancy, ectopic pregnancy, live birth, multiple birth, Cesarean section rates, gestational weeks, maternal obstetric complications, birth weight, newborn sex ratio, and birth defect incidence. Propensity score matching was used to control for potential confounding variables. RESULTS: Of the 2680 cycles included in this study, couples underwent CT examination in 731 cycles. After 1:1 propensity score matching, 670 cycles were included in each group. When comparing demographic and fertility-related variables between groups that had and had not undergone CT examination after propensity score matching, we detected no significant differences in miscarriage rates (16.99% vs. 15.77%, OR = 1.10, 95CI% = 0.74 to 1.68). Similarly, both groups exhibited comparable oocyte and embryonic development, implantation rates (41.99% vs. 40.42%, OR = 1.07, 95%CI = 0.87 to 1.31), clinical pregnancy rates (45.67% vs. 44.48%, OR = 1.05, 95%CI = 0.85 to 1.30), ectopic pregnancy rates (2.94% vs. 1.68%, OR = 1.78, 95%CI = 0.59 to 5.36), live birth rates (36.57% vs. 35.67%, OR = 1.04, 95%CI = 0.83 to 1.30), multiple birth rates, Cesarean section rates, gestational weeks, maternal obstetric complication rates, and neonatal outcomes. CONCLUSIONS: Chest CT examination before IVF/ICSI has no impact on pregnancy and neonatal outcomes associated with fresh embryo transfer. TRIAL REGISTRATION: Not applicable.

Large scale transparency-adjustable mini-LED display with recoverable color gamut by a highly transparent electrochromic shutter.

In this work, a 25 inch (400 × 500 mm) transparency-adjustable mini-LED (TA-MLED) display is constructed of a transparent mini-LED (T-MLED) screen and an electrochromic (EC) shutter. The shutter shows a high transmittance of 86.5% with imperceptible color shift, enabling a perfect vision experience for see-through application. Furthermore, the response speed of the shutter is accelerated by optimal designs in splicing and driving. The coloring time is 55 s, and bleaching time is 36 s. Transmittance of the TA-MLED could be modulated from 3% to 60%. The transparency-adjustable property extends availability of the see-through display screens under strong light irradiations. The T-MLED's color gamut in CIE 1976 shrinks from 145.1% sRGB to 3.6% sRGB with 5161 cd/m2 of backside illumination, and is significantly enhanced to 83.5% sRGB with the active EC shutter.

Transcriptome-wide N6-methyladenine methylation in granulosa cells of women with decreased ovarian reserve.

BACKGROUND: The emerging epitranscriptome plays an essential role in female fertility. As the most prevalent internal mRNA modification, N6-methyladenine (m6A) methylation regulate mRNA fate and translational efficiency. However, whether m6A methylation was involved in the aging-related ovarian reserve decline has not been investigated. Herein, we performed m6A transcriptome-wide profiling in the ovarian granulosa cells of younger women (younger group) and older women (older group). RESULTS: m6A methylation distribution was highly conserved and enriched in the CDS and 3'UTR region. Besides, an increased number of m6A methylated genes were identified in the older group. Bioinformatics analysis indicated that m6A methylated genes were enriched in the FoxO signaling pathway, adherens junction, and regulation of actin cytoskeleton. A total of 435 genes were differently expressed in the older group, moreover, 58 of them were modified by m6A. Several specific genes, including BUB1B, PHC2, TOP2A, DDR2, KLF13, and RYR2 which were differently expressed and modified by m6A, were validated using qRT-PCR and might be involved in the decreased ovarian functions in the aging ovary. CONCLUSIONS: Hence, our finding revealed the transcriptional significance of m6A modifications and provide potential therapeutic targets to promote fertility reservation for aging women.

Endometrial extracellular vesicles of recurrent implantation failure patients inhibit the proliferation, migration, and invasion of HTR8/SVneo cells.

PURPOSE: Endometrial extracellular vesicles are essential in regulating trophoblasts' function. This study aims to investigate whether endometrial extracellular vesicles (EVs) from recurrent implantation failure (RIF) patients inhibit the proliferation, invasion, and migration of HTR8/SVneo cells. METHODS: Eighteen RIF patients and thirteen fertile women were recruited for endometria collection. Endometrial cells isolated from the endometria were cultured and modulated by hormones, and the conditioned medium was used for EV isolation. EVs secreted by the endometrial cells of RIF patients (RIF-EVs) or fertile women (FER-EVs) were determined by Western blotting, nanoparticle tracking analysis, and transmission electron microscopy. Fluorescence-labeled EVs were used to visualize internalization by HTR8/SVneo cells. RIF-EVs and FER-EVs were co-cultured with HTR8/SVneo cells. Cell Counting Kit-8, transwell invasion, and wound closure assays were performed to determine cellular proliferation, invasion, and migration, respectively, in different treatments. RESULTS: RIF-EVs and FER-EVs were bilayer membrane vesicles, ranging from 100 to 150 nm in size, that expressed the classic EV markers Alix and CD9. RIF-EVs and FER-EVs were internalized by HTR8/SVneo cells within 2 h. The proliferation rate in the FER-EV group was significantly higher than that in the RIF-EV group at 20 µg/mL. Moreover, the invasion and migration capacity of trophoblast cells were decreased in the RIF-EV group relative to the FER-EV group at 20 µg/mL. CONCLUSION: Endometrial EVs from RIF patients inhibited the functions of trophoblasts by decreasing their proliferation, migration, and invasive capacity. Such dysregulations induced by RIF-EVs may provide novel insights for better understanding the pathogenesis of implantation failure.

Endometrial extracellular vesicles from women with recurrent implantation failure attenuate the growth and invasion of embryos.

OBJECTIVE: To investigate whether endometrial extracellular vesicles (EVs) from patients with recurrent implantation failure (RIF) attenuate the growth and invasion of embryos. DESIGN: In vitro experimental study. SETTING: University-affiliated hospital. PATIENT(S): Ten RIF patients and seven fertile women. INTERVENTIONS(S): Endometrial cells isolated from endometrial tissues obtained from patients with RIF and fertile women were cultured and modulated in vitro via hormones. Conditioned medium was collected for EV isolation. MAIN OUTCOME MEASURE(S): EVs secreted by endometrial cells of patients with RIF (RIF-EVs) or fertile women (FER-EVs) were characterized with the use of Western blotting, nanoparticle tracking analysis, and transmission electron microscopy. EVs from the two groups were co-cultured with 2-cell murine embryos. Fluorescence-labeled EVs were used to visualize internalization by embryos. Following co-culture, blastocyst and hatching rates were calculated. Blastocysts were stained with diamidino-2-phenylindole to count the total cell number, and the hatched embryos were used to test invasion capacity. RESULT(S): RIF-EVs and FER-EVs are bilayered vesicles ∼100 nm in size and enriched with TSG101, Alix, and CD9. EVs were internalized within 12 hours. The blastocyst rates in the RIF-EV groups were significantly decreased compared with the FER-EV groups at 5, 10, and 20 µg/mL. The hatching rates and total cell numbers of blastocysts also were decreased significantly in the RIF-EV groups compared with the FER-EV groups at 10 and 20 µg/mL. Moreover, the invasion capacity of hatched embryos decreased significantly in the RIF-EV group. CONCLUSION(S): Endometrial EVs from patients with RIF attenuate the development and invasion of embryos.

Aberrant expression of oxytocin receptor in endometrium and decidua in women who have experienced recurrent implantation failure.

OBJECTIVE: To detect the oxytocin receptor (OTR) expression levels in the endometrium and decidua from women who have experienced recurrent implantation failure (RIF) and fertile women. DESIGN: Laboratory study using human endometrial and decidual samples. SETTINGS: University-affiliated hospital. PATIENT(S): Six patients with RIF and six fertile women were recruited for endometrial sampling on day 20-24 of the menstrual cycle. Decidual tissues were collected from women who had a history of RIF and experienced a spontaneous abortion at 6-8 weeks of gestation (n = 8) and women with healthy pregnancies that terminated for nonmedical reasons (n = 8). INTERVENTION: None. MAIN OUTCOME MEASURE(S): OTR expression in the endometrial and decidual tissues was detected with the use of real-time quantitative polymerase chain reaction and Western blotting. RESULT(S): OTR protein and mRNA were significantly increased in the endometria of RIF patients. In the decidua, OTR protein was significantly up-regulated in the RIF group, whereas mRNA was significantly decreased in this group. CONCLUSION(S): Women who experienced RIF presented with an aberrant expression pattern of OTR in the endometria and decidua.