Effect of oral administration of microcin Y on growth performance, intestinal barrier function and gut microbiota of chicks challenged with Salmonella Pullorum.

The lasso peptide microcin Y (MccY) effectively inhibits various serotypes of Salmonella in vitro, but the antibacterial effect against S. Pullorum in poultry is still unclear. This study was the first to evaluate the safety and anti-S. Pullorum infection of MccY in specific pathogen-free (SPF) chicks. The safety test showed that the body weight, IgA and IgM levels of serum, and cecal microbiota structure of 3 groups of chicks orally administrated with different doses of MccY (5 mg/kg, 10 mg/kg, 20 mg/kg) for 14 days were not significantly different from those of the control group. Then, the chicks were randomized into 3 groups for the experiment of anti-S. Pullorum infection: (I) negative control group (NC), (II) S. Pullorum-challenged group (SP, 5 × 108 CFU/bird), (III) MccY-treated group (MccY, 20 mg/kg). The results indicated that compared to the SP group, treatment of MccY increased body weight and average daily gain (P < 0.05), reduced S. Pullorum burden in feces, liver, and cecum (P < 0.05), enhanced the thymus, and decreased the spleen and liver index (P < 0.05). Additionally, MccY increased the jejunal villus height, lowered the jejunal and ileal crypt depth (P < 0.05), and upregulated the expression of IL-4, IL-10, ZO-1 in the jejunum and ileum, as well as CLDN-1 in the jejunum (P < 0.05) compared to the SP group. Furthermore, MccY increased probiotic flora (Barnesiella, etc.), while decreasing (P < 0.05) the relative abundance of pathogenic flora (Escherichia and Salmonella, etc.) compared to the SP group.

International comparison of China's digitalization level and its enlightenment.

The global digital wave has flourished in recent years, and the digital technology revolution has emerged. Digitalization plays an undeniable role in promoting modern economic and social development in multiple aspects such as economy, society, innovation, public services and sustainable development, China's digitalization application is also developing rapidly. In order to better measure the current situation of China's digitalization development level, this paper constructs a comprehensive evaluation index system of digitalization development from four dimensions of talents in the digital field, digital infrastructure construction, digitalization innovation ability and international competitiveness, and tests the index system. The entropy method is used to measure the level of digitalization development between China and the United States, the United Kingdom, France and other major developed countries in the world, and the coefficient of variation method, kernel density estimation and Dagum Gini coefficient method are used to analyze the temporal and spatial characteristics and regional differences of digitalization development level of seven countries. This paper makes a comparative analysis between China and major developed countries from the historical perspective of the evolution of the techno-economic paradigm. With a view to summarizing and exploring from it and drawing on the advanced experiences accumulated by the developed countries over a long period of time, so as to provide China with useful insights and bases for further enhancing its digitalization development level.

Experimental study on the effect of adjacent near-wall heat sources on the particle deposition.

Adjacent near-wall heat sources are widely used in indoor environments. It is important to investigate the particle deposition under the influence of coupled thermal plumes arising from adjacent near-wall heat sources to improve indoor air quality and control harmful particle deposition. Thus, this study scrutinizes the behavior of thermal plumes emanating from adjacent near-wall heat sources, focusing on the deposition of particles with diameters of 0.3 µm, 0.5 µm, 1.0 µm and 3.0 µm on the wall behind the heat sources. These findings are juxtaposed with the pattern of particles with varying sizes situated above the single near-wall heat source and away from the heat sources. The study delves into the impact of varying surface temperatures and the distance from the wall behind the heat sources, as well as the top surface of the heat source, on particle deposition in 29 distinct cases. The results indicate that the deposition velocity of particles with the same size is highest above the adjacent near-wall heat sources, followed by that of a single near-wall heat source, and finally, locations away from the near-wall heat source. Also, the decay rate loss coefficient of particles with the same size above the adjacent near-wall heat sources increases with a decrease in the distance of the heat sources from the wall behind them, an increase in the temperature of the heat sources, and a reduction in distance from the top surface of heat sources.

Influence of PepF peptidase and sporulation on microcin J25 production in Bacillus subtilis.

The lasso peptide microcin J25 (MccJ25) possesses strong antibacterial properties and is considered a potential effective component of bacterial disease treatment drugs and safe food preservatives. Although MccJ25 can be heterologously expressed in Bacillus subtilis as we have previously reported, its regulation and accumulation are yet to be understood. Here, we investigated the expression level and stability of MccJ25 in B. subtilis strains with disruption in peptidase genes pepA, pepF, and pepT. Oligoendopeptidase F (PepF) was found to be involved in reduction of the production of MccJ25 by degradation of its precursor peptide. In the pepF mutant, the MccJ25 reached a concentration of 1.68 µM after a cultivation time exceeding 60 hours, while the wild-type strain exhibited a concentration of only 0.14 µM. Moreover, the production of MccJ25 in B. subtilis downregulated the genes associated with sporulation, and this may contribute to its accumulation. Finally, this study provides a strategy to improve the stability and production of MccJ25 in B. subtilis. IMPORTANCE: MccJ25 displays significant antibacterial activity, a well-defined mode of action, exceptional safety, and remarkable stability. Hence, it presents itself as a compelling candidate for an optimal antibacterial or anti-endotoxin medication. The successful establishment of exogenous production of MccJ25 in Bacillus subtilis provides a strategy for reducing its production cost and diversifying its utilization. In this study, we have provided evidence indicating that both peptidase PepF and sporulation are significant factors that limit the expression of MccJ25 in B. subtilis. The ΔpepF and ΔsigF mutants of B. subtilis express MccJ25 with higher production yield and enhanced stability. To sum up, this study developed several better engineered strains of B. subtilis, which greatly reduced the consumption of MccJ25 during the nutrient depletion stage of the host strain, improved its production, and elucidated factors that may be involved in reducing MccJ25 accumulation in B. subtilis.

Establishing Heterologous Production of Microcins J25 and Y in Bacillus subtilis.

Microcin J25 (MccJ25) and microcin Y (MccY) are lasso peptides and considered potential alternatives to antibiotics and harmful preservatives. The combination of these two microcins can provide a wide antimicrobial spectrum against food-borne Salmonella. Currently, MccJ25 and MccY are produced using Escherichia coli expression systems; however, the entire production process is accompanied by negative effects from endotoxins. In this study, we identified Bacillus subtilis as a suitable host for MccJ25 and MccY production. High-level production of microcins was achieved by promoter optimization, host strain selection, and recombinant expression. The engineered strains produced maximum yields of 2.827 µM MccJ25 and 1.481 µM MccY. This is the first study to demonstrate the expression of MccJ25 and MccY in B. subtilis, and it offers a few engineered strains that are without antibiotic resistance markers, inducer-free, sporulation-deficient, and free of the negative effects of endotoxins for antibacterial therapy and food preservation.

Higher intraoperative mean arterial blood pressure does not reduce postoperative delirium in elderly patients following gastrointestinal surgery: A prospective randomized controlled trial.

BACKGROUND: This study aimed to describe the relationship between the different levels of intraoperative mean arterial blood pressure (MAP) and postoperative delirium in elderly patients undergoing gastrointestinal laparoscopic surgery. MATERIALS AND METHODS: This prospective controlled clinical trial enrolled 116 patients aged 65 to 85 years who underwent gastrointestinal laparoscopic surgery. These patients were randomized 1:1 to a MAP goal of 65 to 85 mmHg (L group) or an 86 to 100 mmHg (H group). The primary endpoint was the incidence of postoperative delirium, assessed twice daily with the Confusion Assessment Method (CAM) and Richmond Agitation-Sedation Scale (RASS) during the first five postoperative days. Delirium severity was evaluated with the Delirium-O-Meter (D-O-M). RESULTS: 108 patients (L group n = 55, H group n = 53) were eventually included in intention-to-treat analyses. Postoperative delirium occurred in 18 (32.7%) of 55 cases of L group and in 15 (28.3%) of 53 cases of H group. The incidence of delirium subtypes between the two groups: hypoactive delirium 14.5% (8/55) vs 11.3% (6/53); hyperactive delirium 7.3% (4/55) vs 3.8% (2/53); mixed delirium 10.9% (6/55) vs 13.2% (7/53). However, the L group showed higher D-O-M scores of the first episode of delirium: 14.5 (Q1 = 12, Q3 = 18.5) vs 12 (Q1 = 10, Q3 = 14), which means the delirium is more severe. CONCLUSIONS: Compared with 65 to 85 mmHg, maintaining intraoperative MAP at 86-100 mmHg did not reduce the incidence of postoperative delirium in elderly patients undergoing gastrointestinal laparoscopic surgery. However, the severity of delirium could be reduced and blood loss is a risk factor for postoperative delirium.

Midazolam exhibits antitumour and enhances the efficiency of Anti-PD-1 immunotherapy in hepatocellular carcinoma.

BACKGROUND: Midazolam (MDZ) is an anaesthetic that is widely used for anxiolysis and sedation. More recently, MDZ has also been described to be related to the outcome of various types of carcinomas. However, how MDZ influences the progression of hepatocellular carcinoma (HCC) and its effects on the biological function and tumour immune microenvironment of this type of tumour remain unknown. METHODS: The effects of MDZ on the proliferation, invasion, and migration of HCC cell lines were examined in vitro using the Cell Counting Kit 8 (CCK8), 5-ethynyl-2'-deoxyuridine (EdU), Transwell, and wound healing assays. Additionally, western blotting was employed to confirm that PD-L1 was expressed. Chromatin immunoprecipitation-seq (ChIP-seq) analysis was used to pinpoint the transcriptional regulation regions of NF-κB and programmed death-ligand 1 (PD-L1). A C57BL/6 mouse model was used to produce subcutaneous HCC tumors in order to evaluate the in vivo performance of MDZ. Mass spectrometry was also used to assess changes in the tumour immunological microenvironment following MDZ injection. RESULTS: The HCC-LM3 and Hep-3B cell lines' proliferation, invasion, and migration were controlled by MDZ, according to the results of the CCK8, EdU, Transwell, and wound healing assays. PD-L1 expression was shown by ChIP-seq analysis to be boosted by NF-κB, and by Western blotting analysis, it was shown that MDZ downregulated the expression of NF-κB. Additionally, in vivo tests revealed that intraperitoneal MDZ injections reduced HCC tumor development and enhanced the effectiveness of anti-PD-1 therapy. The CD45+ immune cell proportions were higher in the MDZ group than in the PBS group, according to the mass spectrometry results. Injection of MDZ resulted in a decrease in the proportions of CD4+ T cells, CD8+ T cells, natural killer (NK) cells, monocytes, Tregs, and M2 macrophages and a rise in the proportion of dendritic cells. Additionally, the concentrations of the cytokines IFN-g and TNF-a were noticeably raised whereas the concentrations of the CD8+ T-cell fatigue markers ICOS, TIGIT, and TIM3 were noticeably lowered. CONCLUSION: According to this study, MDZ inhibited the progression of HCC by inhibiting the NF-κB pathway and reducing the exhaustion of CD8+ T cells. In clinical practice, MDZ combined with anti-PD-1 therapy might contribute to synergistically improving the antitumor efficacy of HCC treatment.

Comparison of preoperative angle kappa measurements in the eyes of cataract patients obtained from Pentacam Scheimpflug system, optical low-coherence reflectometry, and ray-tracing aberrometry.

BACKGROUND: Angle kappa plays a vital role in the implantation of multifocal intraocular lens (MIOL). Large angle kappa is related to a higher risk of postoperative photic phenomena. This study aims to compare preoperative angle kappa in the eyes of cataract patients obtained from the Pentacam Scheimpflug system (Pentacam), optical low-coherence reflectometry (Lenstar), and ray-tracing aberrometry (iTrace). METHODS: One hundred thirteen eyes of 113 patients with cataracts were included. Each eye was examined 3 times using all devices to obtain angle kappa and pupil diameter. When considering dependent eyes for one individual, angle kappa in both right eyes and left eyes should be analysed separately. The repeatability and reproducibility were evaluated using the within-subject standard deviation (Sw), repeatability (2.77 Sw), and intraclass correlation coefficient (ICC). The difference, correlation, and agreement between devices were evaluated by paired t-tests, Pearson tests, and Bland-Altman analysis, respectively. RESULTS: Intraoperator repeatability and interoperator and intersession reproducibility of angle kappa showed an Sw of less than 0.05 mm, a 2.77 Sw of 0.14 mm or less, and an ICC of more than 0.96. Angle kappa was not significantly different between Pentacam and Lenstar (P > 0.05), while angle kappa was significantly different between Pentacam and iTrace and between Lenstar and iTrace (P < 0.05). There was a strong correlation between Pentacam and Lenstar for angle kappa (r =0.907 to 0.918) and a weak or moderate correlation between Pentacam and iTrace and between Lenstar and iTrace (r =0.292 to 0.618). There were narrow 95% limits of agreement (LoA) between Pentacam and Lenstar for angle kappa and wide 95% LoA between Pentacam and iTrace and between Lenstar and iTrace. No significant differences in pupil diameter were found between Pentacam and Lenstar in either eye (P > 0.05). Positive angle kappa (nasal light reflex) was found in most cataract patients (79.25% to 84.91%) through 3 different devices in both eyes. CONCLUSIONS: The 3 devices provided high intraoperator repeatability and interoperator and intersession reproducibility for angle kappa measurements. The measurement of preoperative angle kappa in the eyes of patients with cataracts by Pentacam and Lenstar has good agreement.

SYVN1-mediated ubiquitination and degradation of MSH3 promotes the apoptosis of lens epithelial cells.

The pathology of age-related cataract (ARC) mainly involves the misfolding and aggregation of proteins, especially oxidative damage repair proteins, in the lens, induced by ultraviolet-B (UVB). MSH3, as a key member of the mismatch repair family, primarily maintains genome stability. However, the function of MSH3 and the mechanism by which cells maintain MSH3 proteostasis during cataractogenesis remains unknown. In the present study, the protein expression levels of MSH3 were found to be attenuated in ARC specimens and SRA01/04 cells under UVB exposure. The ectopic expression of MSH3 notably impeded UVB-induced apoptosis, whereas the knockdown of MSH3 promoted apoptosis. Protein half-life assay revealed that UVB irradiation accelerated the decline of MSH3 by ubiquitination and degradation. Subsequently, we found that E3 ubiquitin ligase synoviolin (SYVN1) interacted with MSH3 and promoted its ubiquitination and degradation. Of note, the expression and function of SYVN1 were contrary to those of MSH3 and SYVN1 regulated MSH3 protein degradation via the ubiquitin-proteasome pathway and the autophagy-lysosome pathway. Based on these findings, we propose a mechanism for ARC pathogenesis that involves SYVN1-mediated degradation of MSH3 via the ubiquitin-proteasome pathway and the autophagy-lysosome pathway, and suggest that interventions targeting SYVN1 might be a potential therapeutic strategy for ARC.

Insights into glucosinolate accumulation and metabolic pathways in Isatis indigotica Fort.

BACKGROUND: Glucosinolates (GSLs) play important roles in defending against exogenous damage and regulating physiological activities in plants. However, GSL accumulation patterns and molecular regulation mechanisms are largely unknown in Isatis indigotica Fort. RESULTS: Ten GSLs were identified in I. indigotica, and the dominant GSLs were epiprogoitrin (EPI) and indole-3-methyl GSL (I3M), followed by progoitrin (PRO) and gluconapin (GNA). The total GSL content was highest (over 20 µmol/g) in reproductive organs, lowest (less than 1.0 µmol/g) in mature organs, and medium in fresh leaves (2.6 µmol/g) and stems (1.5 µmol/g). In the seed germination process, the total GSL content decreased from 27.2 µmol/g (of seeds) to 2.7 µmol/g (on the 120th day) and then increased to 4.0 µmol/g (180th day). However, the content of indole GSL increased rapidly in the first week after germination and fluctuated between 1.13 µmol/g (28th day) and 2.82 µmol/g (150th day). Under the different elicitor treatments, the total GSL content increased significantly, ranging from 2.9-fold (mechanical damage, 3 h) to 10.7-fold (MeJA, 6 h). Moreover, 132 genes were involved in GSL metabolic pathways. Among them, no homologs of AtCYP79F2 and AtMAM3 were identified, leading to a distinctive GSL profile in I. indigotica. Furthermore, most genes involved in the GSL metabolic pathway were derived from tandem duplication, followed by dispersed duplication and segmental duplication. Purifying selection was observed, although some genes underwent relaxed selection. In addition, three tandem-arrayed GSL-OH genes showed different expression patterns, suggesting possible subfunctionalization during evolution. CONCLUSIONS: Ten different GSLs with their accumulation patterns and 132 genes involved in the GSL metabolic pathway were explored, which laid a foundation for the study of GSL metabolism and regulatory mechanisms in I. indigotica.

Influence of angle alpha on visual quality after implantation of extended depth of focus intraocular lenses.

BACKGROUND: To assess postoperative changes in angle alpha, and to evaluate the postoperative visual quality of patients with different angle alpha values after implantation of extended depth of focus (EDOF) intraocular lenses (IOLs). METHODS: Seventy-nine eyes of 79 patients who had phacoemulsification with EDOF IOLs implantation were enrolled. A cut-off value of 0.3 mm, 0.4 mm, and 0.5 mm in preoperative angle alpha was chosen to divide eyes into groups. Distance, intermediate, and near visual acuities, modulation transfer function (MTF), and aberrations were recorded during a 6-month follow-up. A patient questionnaire was completed. RESULTS: There were no significant differences in angle alpha postoperatively compared to preoperatively. No significant differences were found in visual acuity and MTF between all groups. With 5 mm pupil diameter, there were significant differences of higher-order aberrations and spherical aberration in ocular aberration and internal aberration between angle alpha<0.4 mm and angle alpha≥0.4 mm. Additionally, significant differences of coma were also added in cut-off value of 0.5 mm. When the value of angle alpha is 0.4 mm or higher, there were significant differences in the score of halos and glare. CONCLUSIONS: Angle alpha did not affect visual acuity, but the value of 0.4 mm or higher in angle alpha affected the visual quality under scotopic conditions and occurrence of halos and glare. For patients with 0.4 mm or higher in angle alpha, the choice to implant a EDOF IOL should be carefully considered.

Cytokines possibly involved in idiopathic epiretinal membrane progression after uncomplicated cataract surgery.

Epiretinal membrane (ERM) is a common retinal fibrotic disorder disease causing visual impairment and metamorphopsia. Recently, increasing attention has been devoted to ERM progression after uncomplicated cataract surgery. Cytokines, which play a role in diverse physiological and pathological activities in eyes, are suggested to be involved in these postoperative changes. However, few studies have investigated the post-cataract surgery cytokine expression changes in ERM eyes and their roles in the postoperative changes. The purpose of this study was to evaluate the aqueous levels of cytokines in eyes with idiopathic epiretinal membrane (iERM) both pre- and post-cataract surgery, and their correlations with postoperative iERM progression. In this study, aqueous humor (AH) samples were collected from iERM eyes (n = 25) and non-iERM eyes (n = 23) from 48 patients (48 eyes) undergoing uncomplicated cataract surgery preoperatively and 20 h postoperatively. Samples were analyzed for 48 cytokines with multiplex bead-based immunoassay. Correlations between cytokine level changes (postoperation vs. preoperation) and three-month postoperative best-corrected visual acuity (BCVA) and optical coherence tomography measure changes were evaluated in iERM eyes. We found that in iERM eyes, the levels of 4 cytokines exhibited significant elevations when compared with those in the controls (all p ≤ 0.0015) preoperatively. Postoperatively, the concentrations of 21 cytokines were higher than the preoperative levels in iERM eyes (all p ≤ 0.0015), among which GRO-α, IL-8, and MCP-3 levels showed more pronounced changes than the controls. Additionally, in iERM eyes, IL-4 level changes showed moderate positive correlations with MV (r = 0.492, p = 0.028) and MT (r = 0.481, p = 0.032) changes. LogMAR changes were positively correlated with IL-1α (r = 0.553, p = 0.011), IL-12(P40) (r = 0.544, p = 0.013), and MCP-3 (r = 0.588, p = 0.006) level changes. No significant cytokine-level-change differences were found between eyes with and without postoperative cystoid macular edema development. In conclusion, cataract surgery will bring great alterations to the specific intraocular cytokine microenvironment inherently in eyes with iERM. Many fibrotic and inflammatory cytokines showing elevated levels or relationships with clinical characteristics are suggested to be involved in the pathogenesis and post-cataract surgery progression of iERM; however, further investigations are needed to discern their real roles.

Current Perspectives on B Lymphocytes in the Immunobiology of Hepatocellular Carcinoma.

Immune cells infiltrating tumors are capable of significantly impacting carcinogenesis through cancer promotion and anticancer responses. There are many aspects of hepatocellular carcinoma (HCC) related T lymphocytes that are undergoing extensive studies, whereas the effect exerted by B lymphocytes remains a less researched area. In this study, the latest research on the effect of B lymphocytes as they infiltrate tumors in relation to HCC is presented. Their prognosis-related importance is analyzed, along with their function in the tumor microenvironment (TME), as well as the way that B cell biology can be employed to help create a B cell therapy strategy for HCC.

Profiling and Integrated Analysis of the ERCC6-regulated circRNA-miRNA-mRNA Network in Lens Epithelial Cells.

Purpose: To explore the regulatory role of ERCC6 in the circRNA-miRNA-mRNA network using a cellular ERCC6 overexpression model (OE-ERCC6) in lens epithelial cells.Methods: The expression profiles of circRNAs, miRNAs and mRNAs were determined by RNA-seq, and a regulatory circRNA-miRNA-mRNA network was constructed via bioinformatics. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were used for the functional annotation of circRNA host genes, differentially expressed (DE) genes, and miRNA targets.Results: The DE molecules between the OE-ERCC6 and control groups included 269 circRNAs, 241 miRNAs and 3500 mRNAs. We validated 5 selected DE reads of circRNAs (hsa_circ_0001009, hsa_circ_0002024, hsa_circ_0004592, hsa_circ_0001900 and hsa_circ_0001017). Subsequent bioinformatics analysis revealed that the DE circRNAs are mainly involved in oxidative stress- and cell death-related signaling pathways. Finally, a circRNA-miRNA-mRNA network focusing on DNA damage and cell death, which involved 5 circRNAs, 13 miRNAs and 107 mRNAs, was constructed.Conclusion: We constructed a circRNA-miRNA-mRNA network that is regulated by ERCC6. DE circRNAs have the potential to become therapeutic targets related to the lens lesions observed in ARC. The establishment of related in vivo and in vitro models could be a future direction to confirm these hypotheses.

Comprehensive Profiling of Proteome and Ubiquitome Changes in Human Lens Epithelial Cell Line after Ultraviolet-B Irradiation.

Ultraviolet-B (UVB) is a recognized risk factor for age-related cataract (ARC) and can cause various changes, including ubiquitination, in lens epithelial cells (LECs). However, the relationship between ubiquitination and ARC is unclear. Herein, we used UVB-irradiated human lens epithelial cell line (SRA01/04) representing the cell model of ARC to investigate the profile changes in the proteome and ubiquitome. A total of 552 differentially expressed proteins (DEPs) and 871 differentially ubiquitinated proteins (DUPs) were identified, and 9 ubiquitination motifs were found. Bioinformatics analysis revealed diverse pathways and biological processes of differential proteins and several DNA damage repair proteins that were potentially mediated via ubiquitin-proteasome pathway. We validated the decreased protein expression of DNA-directed RNA polymerase II subunit RPB2 (POLR2B) in both human cataract capsule tissues and UVB-treated SRA01/04 cells and found that treatment with proteasome inhibitor (MG-132) could reverse the protein level of POLR2B in UVB-irradiated SRA01/04 cells. Our data provide novel information regarding protein expressions and ubiquitination modifications in UVB-induced oxidative damage model. This study might offer a cell-level reference to further investigate the pathogenesis of ARC.

Seed-Specific Overexpression of SPL12 and IPA1 Improves Seed Dormancy and Grain Size in Rice.

Pre-harvest sprouting (PHS) often results in reduced grain yield and quality and is a major problem in cereal production. Improved seed dormancy would inhibit PHS. Here we show that seed-specific overexpression of two SQUAMOSA-PROMOTER BINDING PROTEIN-LIKE (SPL) genes SPL12 and IPA1 enhances seed dormancy and inhibits PHS without noticeable effects on shoot architecture in rice. In addition, seed-specific overexpression of IPA1 also increases grain size and thus improves grain productivity. Furthermore, our results suggest that SPL12 enhances the seed dormancy through directly regulating many genes in the gibberellin (GA) pathway. This research provides an efficient method to suppress PHS and will facilitate breeding elite crop varieties.

Identification and Characterization of N6-Methyladenosine CircRNAs and Methyltransferases in the Lens Epithelium Cells From Age-Related Cataract.

Purpose: To explore the involvement of N6-methyladenosine (m6A) modification in circular RNAs (circRNAs) and relevant methyltransferases in the lesion of lens epithelium cells (LECs) under the circumstances of age-related cataract (ARC). Methods: LECs were collected from normal subjects and patients with cortical type of ARC (ARCC). M6A-tagged circRNAs and circRNAs expression were analyzed by m6A-modified RNA immunoprecipitation sequencing (m6A-RIP-seq) and RNA sequencing (RNA-seq). Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were used to predict possible functions of the m6A-circRNAs. Expression of m6A-related methyltransferase and demethytransferase was measured by quantitative real-time polymerase chain reaction. Expression and location of AlkB homolog 5 RNA demethylase (ALKBH5), a key component of m6A demethytransferase, were determined by Western blot and immunostaining. Results: All 4646 m6A peaks within circRNAs had different abundances, with 2472 enriched and 2174 subdued. The level of m6A abundance in total circRNAs was decreased in the LECs from ARCCs in comparison with the controls. We also found that the expression of highly m6A-tagged circRNAs was mostly decreased in comparison with non-m6A-tagged circRNAs. The bioinformatics analysis predicted the potential functions of m6A modified circRNAs and the relevant pathways that may be associated with m6A modified circRNAs. Among five major methyltransferases, ALKBH5 was significantly upregulated in LECs of ARCCs. Conclusions: Our data provided novel evidence regarding the involvement of circRNAs m6A modifications in ARC. The altered expression of methyltransferases in lens tissue might selectively change the epigenetic profile of lens genome through regulating genes that host the circRNAs, thus enhance the susceptibility to ARC. The results might provide a new insight in the molecular target of ARC pathogenesis.

Genome-Wide Identification and Analysis on YUCCA Gene Family in Isatis indigotica Fort. and IiYUCCA6-1 Functional Exploration.

Auxin is one of the most critical hormones in plants. YUCCA (Tryptophan aminotransferase of Arabidopsis (TAA)/YUCCA) enzymes catalyze the key rate-limiting step of the tryptophan-dependent auxin biosynthesis pathway, from IPA (Indole-3-pyruvateacid) to IAA (Indole-3-acetic acid). Here, 13 YUCCA family genes were identified from Isatis indigotica, which were divided into four categories, distributing randomly on chromosomes (2n = 14). The typical and conservative motifs, including the flavin adenine dinucleotide (FAD)-binding motif and flavin-containing monooxygenases (FMO)-identifying sequence, existed in the gene structures. IiYUCCA genes were expressed differently in different organs (roots, stems, leaves, buds, flowers, and siliques) and developmental periods (7, 21, 60, and 150 days after germination). Taking IiYUCCA6-1 as an example, the YUCCA genes functions were discussed. The results showed that IiYUCCA6-1 was sensitive to PEG (polyethylene glycol), cold, wounding, and NaCl treatments. The over-expressed tobacco plants exhibited high auxin performances, and some early auxin response genes (NbIAA8, NbIAA16, NbGH3.1, and NbGH3.6) were upregulated with increased IAA content. In the dark, the contents of total chlorophyll and hydrogen peroxide in the transgenic lines were significantly lower than in the control group, with NbSAG12 downregulated and some delayed leaf senescence characteristics, which delayed the senescence process to a certain extent. The findings provide comprehensive insight into the phylogenetic relationships, chromosomal distributions, and expression patterns and functions of the YUCCA gene family in I. indigotica.

Comparison of optical quality and distinct macular thickness in femtosecond laser-assisted versus phacoemulsification cataract surgery.

BACKGROUND: Optical quality and macular thickness changing optical quality is rarely reported after femtosecond laser-assisted cataract surgery (FLACS). In current research, we evaluated optical quality recovery and distinct macular thickness changes after FLACS and phacoemulsification cataract surgery (PCS). METHODS: A total of 100 cataract patients (100 eyes) were included (50 eyes for the FLACS group and 50 eyes for the PCS group). Modulation transfer function (MTF), point spread function (PSF) and dysfunctional lens index (DLI) were measured by a ray-tracing aberrometer (iTrace). Uncorrected distance visual acuity (UDVA) and corrected distance visual acuity (CDVA) were also assessed pre-operation,1 week and 1 month after surgery. The MTF values at spatial frequencies of 5, 10, 15, 20, 25 and 30 cycles/degree (c/d) were selected. We used optical coherence tomography (OCT) to assess the macular thickness of different regions pre-operatively and1month after the surgery. RESULTS: In PCS group, we found the statistically significant differences between pre-operation and post-operation in DLI (p < 0.0001), PSF (strehl ratio, SR) (p = 0.027) and MTF (p = 0.028), but not intraocular pressure (IOP) (p = 0.857). The differences between pre-operation and post-operation for DLI (p = 0.031), SR (p = 0.01) and IOP (p = 0.03), but not MTF (p = 0.128) were also found in FLACS group. The differences were statistically significant when the spatial frequencies were at 5, 10 and 25 (p = 0.013, 0.031 and 0.048) between pre-operation and post-operation in PCS group but not FLACS group at 1 month. In PCS group, we found the differences between pre-operation and post-operation in nasal inter macular ring thickness (NIMRT) (p = 0.03), foveal volume (FV) (p = 0.034) and average retinal thickness (ART) (p = 0.025) but not FLACS group at 1 month. CONCLUSION: FLACS is safe that did not cause significant increase of macular thickness in current study. However, it also cannot produce better optical quality. In contrast, PCS can produce macular thickness changes, but better optical quality recovery. The slightly retinal change may not affect optical quality.