FTO-mediated DSP m6A demethylation promotes an aggressive subtype of growth hormone-secreting pituitary neuroendocrine tumors.

BACKGROUND: Growth hormone-secreting pituitary neuroendocrine tumors can be pathologically classified into densely granulated (DGGH) and sparsely granulated types (SGGH). SGGH is more aggressive and associated with a poorer prognosis. While epigenetic regulation is vital in tumorigenesis and progression, the role of N6-methyladenosine (m6A) in aggressive behavior has yet to be elucidated. METHODS: We performed m6A-sequencing on tumor samples from 8 DGGH and 8 SGGH patients, complemented by a suite of assays including ELISA, immuno-histochemistry, -blotting and -fluorescence, qPCR, MeRIP, RIP, and RNA stability experiments, aiming to delineate the influence of m6A on tumor behavior. We further assessed the therapeutic potential of targeted drugs using cell cultures, organoid models, and animal studies. RESULTS: We discovered a significant reduction of m6A levels in SGGH compared to DGGH, with an elevated expression of fat mass and obesity-associated protein (FTO), an m6A demethylase, in SGGH subtype. Series of in vivo and in vitro experiments demonstrated that FTO inhibition in tumor cells robustly diminishes hypoxia resistance, attenuates growth hormone secretion, and augments responsiveness to octreotide. Mechanically, FTO-mediated m6A demethylation destabilizes desmoplakin (DSP) mRNA, mediated by the m6A reader FMR1, leading to prohibited desmosome integrity and enhanced tumor hypoxia tolerance. Targeting the FTO-DSP-SSTR2 axis curtailed growth hormone secretion, therefor sensitizing tumors to octreotide therapy. CONCLUSION: Our study reveals the critical role of FTO in the aggressive growth hormone-secreting pituitary neuroendocrine tumors subtype and suggests FTO may represent a new therapeutic target for refractory/persistent SGGH.

High correlated color temperature artificial lighting impairs retinal pigment epithelium integrity and chloride ion transport: A potential mechanism for choroidal thinning.

Among the environmental factors contributing to myopia, the role of correlated color temperature (CCT) of ambient light emerges as a key element warranting in-depth investigation. The choroid, a highly vascularized and dynamic structure, often undergoes thinning during the progression of myopia, though the precise mechanism remains elusive. The retinal pigment epithelium (RPE), the outermost layer of the retina, plays a pivotal role in regulating the transport of ion and fluid between the subretinal space and the choroid. A hypothesis suggests that variations in choroidal thickness (ChT) may be modulated by transepithelial fluid movement across the RPE. Our experimental results demonstrate that high CCT illumination significantly compromised the integrity of tight junctions in the RPE and disrupted chloride ion transport. This functional impairment of the RPE may lead to a reduction in fluid transfer across the RPE, consequently resulting in choroidal thinning and potentially accelerating axial elongation. Our findings provide support for the crucial role of the RPE in regulating ChT. Furthermore, we emphasize the potential hazards posed by high CCT artificial illumination on the RPE, the choroid, and refractive development, underscoring the importance of developing eye-friendly artificial light sources to aid in the prevention and control of myopia.

Dual single guide RNAs-mediating deletion of mature myostatin peptide results in concomitant muscle fibre hyperplasia and adipocyte hypotrophy in pigs.

Myostatin (MSTN) is a major gene target for skeletal muscle overgrowth in animals. We hypothesized that deletion of the entire mature peptide encoded by MSTN in pigs would knock out its bioactive form and accordingly stimulate skeletal muscle overgrowth. Thus, we engineered two pairs of single-guide RNAs (sgRNAs) to target exons 1 and 3 of MSTN in primary fetal fibroblasts of Taoyuan black pigs. We found that sgRNAs targeting exon 3, which encodes the mature peptide, had higher biallelic null mutation efficiency than those targeting exon 1. Somatic cell nuclear transfer was conducted using the exon 3 mutation cells as donor cells to generate five cloned MSTN null piglets (MSTN-/-). Growth testing revealed that both the growth rate and average daily weight gain of MST-/- pigs were greater than those of wild-type (MSTN+/+) pigs. Slaughter data demonstrated that the lean ratio of MSTN-/- pigs was 11.3% higher (P < 0.01) while the back-fat thickness was 17.33% lower (P < 0.01) than those of MSTN+/+ pigs. Haematoxylin-eosin staining indicated that the increased leanness of MSTN-/- pigs resulted from muscle fibre hyperplasia rather than hypertrophy.HE staining showed markedly decreased adipocyte size in MSTN-/- pigs. We also critically examined the off-target and random integration by resequencing, which showed that the founder MSTN-/- pigs contained no non-target mutations or exogenous plasmid elements. This study is the first to report the successful knock out of the mature MSTN peptide using dual sgRNA-mediated deletion, leading to the most prominent alteration of meat production traits in pigs published thus far. This new strategy is expected to have a wide impact on genetic improvements in food animals.

Mapping and candidate gene analysis of clustered bud on the main inflorescence in Brassica napus L.

Breeding rapeseed varieties with more main inflorescence siliques is an idea for developing rapeseed varieties that are suitable for light and simplified cultivation. The Brassica napus exhibited cluster bud of the main inflorescence (Bnclib) gene. At the fruiting stage, the main inflorescence had more siliques, higher density, and more main inflorescences. Moreover, the top of the main inflorescence bifurcated. Genetic analysis showed that the separation ratio between Bnclib and the wild type in the F2 generation was 3:1, which indicated that the trait was a single-gene-dominant inheritance. Among the 24 candidate genes, only one gene, BnaA03g53930D, showed differential expression between the groups (False discovery rate, FDR ≤ 0.05, |log2FC|≤ 1). qPCR verification of the BnaA03g53930D gene between Huyou 17 and its Bnclib near-isogenic line showed that BnaA03g53930D was significantly differentially expressed in the stem tissue of Huyou 17 and its Bnclib near-isogenic line (Bnclib NIL). The determination of gibberellin (GA), brassinolide (BR), cytokinin (CTK), jasmonic acid (JA), growth hormone (IAA), and strigolactone (SL) content in the shoot apex of Huyou 17 by Bnclib NIL and wild type showed that all six hormones significantly differed between the Bnclib NIL and Huyou 17. It is necessary to conduct further research on the interactions between JA and the other five hormones and the main inflorescence bud clustering in B. napus.

Deeply Reconstructed Hierarchical Ni-Co Microwire for Flexible Ni-Zn Microbattery with Excellent Comprehensive Performance.

The rise of flexible electronics calls for efficient microbatteries (MBs) with requirements in energy/power density, stability, and flexibility simultaneously. However, the ever-reported flexible MBs only display progress around certain aspects of energy loading, reaction rate, and electrochemical stability, and it remains challenging to develop a micro-power source with excellent comprehensive performance. Herein, a reconstructed hierarchical Ni-Co alloy microwire is designed to construct flexible Ni-Zn MB. Notably, the interwoven microwires network is directly formed during the synthesis process, and can be utilized as a potential microelectrode which well avoids the toxic additives and the tedious traditional powder process, thus greatly simplifying the manufacture of MB. Meanwhile, the hierarchical alloy microwire is composed of spiny nanostructures and highly active alloy sites, which contributes to deep reconstruction (≈100 nm). Benefiting from the dense self-assembled structure, the fabricated Ni-Zn MB obtained high volumetric/areal energy density (419.7 mWh cm-3 , 1.3 mWh cm-2 ), and ultrahigh rate performance extending the power density to 109.4 W cm-3 (328.3 mW cm-2 ). More surprisingly, the MB assembled by this inherently flexible microwire network is extremely resistant to bending/twisting. Therefore, this novel concept of excellent comprehensive micro-power source will greatly hold great implications for next-generation flexible electronics.

Enhanced Formation of 6PPD-Q during the Aging of Tire Wear Particles in Anaerobic Flooded Soils: The Role of Iron Reduction and Environmentally Persistent Free Radicals.

Rapid urbanization drives increased emission of tire wear particles (TWPs) and the contamination of a transformation product derived from tire antioxidant, termed as N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine-quinone (6PPD-Q), with adverse implications for terrestrial ecosystems and human health. However, whether and how 6PPD-Q could be formed during the aging of TWPs in soils remains poorly understood. Here, we examine the accumulation and formation mechanisms of 6PPD-Q during the aging of TWPs in soils. Our results showed that biodegradation predominated the fate of 6PPD-Q in soils, whereas anaerobic flooded conditions were conducive to the 6PPD-Q formation and thus resulted in a ∼3.8-fold higher accumulation of 6PPD-Q in flooded soils than wet soils after aging of 60 days. The 6PPD-Q formation in flooded soils was enhanced by Fe reduction-coupled 6PPD oxidation in the first 30 days, while the transformation of TWP-harbored environmentally persistent free radicals (EPFRs) to superoxide radicals (O2•-) under anaerobic flooded conditions further dominated the formation of 6PPD-Q in the next 30 days. This study provides significant insight into understanding the aging behavior of TWPs and highlights an urgent need to assess the ecological risk of 6PPD-Q in soils.

Effect of Wearing N95 Mask on the Quality of Chest Compressions in Prehospital Emergency Personnel: A Cross-over Study.

OBJECTIVE: To evaluate the effect of wearing an N95 mask on the quality of chest compression and fatigue of prehospital emergency personnel during cardiopulmonary resuscitation (CPR). METHODS: Twenty-four eligible participants were recruited. Participants' age, sex, height, and weight were recorded. After completing the CPR training and examination, participants were tested twice, wearing surgical mask or an N95 mask, while performing chest compressions for 2 minutes. The quality of chest compression (including compression frequency, depth, rebound, and position) was recorded by the simulator. Borg fatigue scores and physiological parameters (including heart rate, mean arterial pressure, pulse oxygen saturation, and respiratory rate) were recorded before and after chest compressions. RESULTS: Compared to wearing surgical masks, participants wearing N95 masks had significantly lower quality of chest compression, including compression frequency (98.3 ± 4.9 bpm vs 104.0 ± 6.0 bpm, P < 0.001), depth (47.1 ± 4.5 mm vs 50.5 ± 5.4 mm, P < 0.001), and rebound (90.2 ± 2.7% vs 94.3 ± 2.1%, P < 0.001). The compression position was not affected. The period data showed that the difference in compression quality started after 1 minute of compressions. Participants wearing N95 masks had higher Borg fatigue scores [6.1(2) vs 5.1(2), P < 0.001], heart rates (121.2 ± 5.7 bpm vs 109.9 ± 6.0 bpm, P < 0.001), mean arterial pressures (106.3 ± 8.0 mmHg vs 99.0 ± 8.5 mmHg, P = 0.012), and respiratory rates (29.5 ± 2.7 bpm vs 24.7 ± 2.5 bpm, P < 0.001). CONCLUSION: This study showed that the use of an N95 mask by prehospital emergency personnel during the performance of chest compressions resulted in a reduction of compression quality and increased clinician fatigue. There is a need for CPR training for medical personnel wearing personal protective equipment.

Application of a preoperative pain management mode based on instant messaging software in elderly hip fracture patients: a randomized controlled trial.

BACKGROUND: Preoperative analgesia of hip fracture in elderly patients is important, but it is also lacking. In particular, nerve block was not provided in time. In order to provide more effective analgesia, we designed a multimodal pain management mode based on instant messaging software. METHODS: From May to September 2022, a total of 100 patients with unilateral hip fracture aged over 65 were randomly divided into the test group and the control group. Finally, 44 patients in each group completed the result analysis. A new pain management mode was used in the test group. This mode focuses on the full information exchange between medical personnel in different departments, early fascia iliaca compartment block (FICB), and closed-loop pain management. Outcomes include the time when FICB is completed for the first time; The number of cases of FICB completed by emergency doctors; Patients' pain score, pain duration. RESULTS: The time for patients in the test group to complete FICB for the first time was 3.0 [1.925-3.475] h, which was less than the time for patients in the control group (4.0 [3.300-5.275] h). The difference was statistically significant (P < 0.001). Compared with 16 patients in the control group, 24 patients in the test group completed FICB by emergency doctors, and there was no statistical difference between the two groups (P = 0.087). The test group was superior to the control group in the highest NRS score (4.00 [3.00-4.00] vs 5.00 [4.00-5.75]), the duration of the highest NRS score (20.00 [20.00-25.00] mins vs 40.00 [30.00-48.75] mins), and the NRS > 3 time (35.00 [20.00-45.00] mins vs 72.50 [60.00-45.00] mins). The analgesic satisfaction of patients in the test group (5.00 [4.00-5.00]) was also significantly higher than that of the control group (3.00 [3.00-4.00]). The above four indexes were different between the two groups (P < 0.001). CONCLUSIONS: Using instant messaging software, the new model of pain management can enable patients to receive FICB as soon as possible and improve the timeliness and effectiveness of analgesia. TRIAL REGISTRATION: Chinese Clinical Registry Center, ChiCTR2200059013, 23/04/2022.

Construction and validation of a risk prediction model for intraoperative hypothermia in elderly patients undergoing total hip arthroplasty.

AIMS: To construct and validate an intraoperative hypothermia risk prediction model for elderly patients undergoing total hip arthroplasty (THA). METHODS: We collected data from 718 patients undergoing THA in a tertiary hospital from January 2021 to December 2022. Of these patients, 512 were assigned to the modeling group from January 2021 to April 2022, and 206 participants were assigned to the validation group from May 2022 to December 2022. A logistic regression analysis was performed to construct the model. The area under the curve (AUC) was used to test the model's predictive ability. RESULTS: The incidence rate of intraoperative hypothermia was 51.67%. The risk factors entered into the risk prediction model were age, preoperative hemoglobin level, intraoperative blood loss, postoperative hemoglobin level, and postoperative systolic blood pressure. The model was constructed as follows: logit (P) = - 10.118 + 0.174 × age + 1.366 × 1 (preoperative hemoglobin level) + 0.555 × 1 (postoperative hemoglobin level) + 0.009 × 1 (intraoperative blood loss) + 0.066 × 1 (postoperative systolic blood pressure). Using the Hosmer-Lemeshow test, the P value was 0.676 (AUC, 0.867). The Youden index, sensitivity, and specificity were 0.602, 0.790, and 0.812, respectively. The incidence rates of intraoperative hypothermia in the modeling and validation groups were 53.15% and 48.06%, respectively. The correct practical application rate was 89.81%. This model had good application potential. CONCLUSIONS: This risk prediction model has good predictive value and can accurately predict the occurrence of intraoperative hypothermia in patients who undergo THA, which provides reliable guidance for clinical work and has good clinical application value.

TET Family Members Are Integral to Porcine Oocyte Maturation and Parthenogenetic Pre-Implantation Embryogenesis.

The ten-eleven translocation (TET) enzyme family, which includes TET1/2/3, participates in active DNA demethylation in the eukaryotic genome; moreover, TET1/2/3 are functionally redundant in mice embryos. However, the combined effect of TET1/2/3 triple-gene knockdown or knockout on the porcine oocytes or embryos is still unclear. In this study, using Bobcat339, a specific small-molecule inhibitor of the TET family, we explored the effects of TET enzymes on oocyte maturation and early embryogenesis in pigs. Our results revealed that Bobcat339 treatment blocked porcine oocyte maturation and triggered early apoptosis. Furthermore, in the Bobcat339-treated oocytes, spindle architecture and chromosome alignment were disrupted, probably due to the huge loss of 5-hydroxymethylcytosine (5hmC)and concurrent increase in 5-methylcytosine (5mC). After Bobcat339 treatment, early parthenogenetic embryos exhibited abnormal 5mC and 5hmC levels, which resulted in compromised cleavage and blastocyst rate. The mRNA levels of EIF1A and DPPA2 (ZGA marker genes) were significantly decreased, which may explain why the embryos were arrested at the 4-cell stage after Bobcat339 treatment. In addition, the mRNA levels of pluripotency-related genes OCT4 and NANOG were declined after Bobcat339 treatment. RNA sequencing analysis revealed differentially expressed genes in Bobcat339-treated embryos at the 4-cell stage, which were significantly enriched in cell proliferation, cell component related to mitochondrion, and cell adhesion molecule binding. Our results indicated that TET proteins are essential for porcine oocyte maturation and early embryogenesis, and they act by mediating 5mC/5hmC levels and gene transcription.