Acrolein Induces Retinal Abnormalities of Alzheimer's Disease in Mice.

It is reported that retinal abnormities are related to Alzheimer's disease (AD) in patients and animal models. However, it is unclear whether the retinal abnormities appear in the mouse model of sporadic Alzheimer's disease (sAD) induced by acrolein. We investigated the alterations of retinal function and structure, the levels of ß-amyloid (Aß) and phosphorylated Tau (p-Tau) in the retina, and the changes in the retinal vascular system in this mouse model. We demonstrated that the levels of Aß and p-Tau were increased in the retinas of mice from the acrolein groups. Subsequently, a decreased amplitudes of b-waves in the scotopic and photopic electroretinogram (ERG), decreased thicknesses of the retinal nerve fiber layer (RNFL) in the retina, and slight retinal venous beading were found in the mice induced by acrolein. We propose that sAD mice induced by acrolein showed abnormalities in the retina, which may provide a valuable reference for the study of the retina in sAD.

Effects of PPAR-γ and RXR-α on mouse meibomian gland epithelial cells during inflammation induced by latanoprost.

The purpose of this study is to investigate the effects of latanoprost on the secretion of cytokines and chemokines from meibomian gland epithelial cells, and to evaluate the modulation of peroxisome proliferator-activated receptor γ (PPAR-γ) and retinoid X receptor α (RXR-α) during latanoprost-induced inflammation. Mouse meibomian gland epithelial cells were cultured in proliferation and differentiation medium, respectively. Cells were exposed to latanoprost, rosiglitazone (PPAR-γ agonist), or LG100268 (RXR-α agonist), respectively. The expression of IL-6, IL-1ß, TNF-α, MMP-9, MCP-1, and CCL-5 were detected by real-time PCR and ELISA. The effect of latanoprost, rosiglitazone, LG100268, and inflammatory cytokines on the differentiation of meibocyte were evaluated by related gene expression and lipid staining. The expression of Keratin-1, 6, 17 protein was detected by western immunoblotting. The results showed that the above cytokines could be induced by latanoprost in meibomian gland epithelial cells. LG100268 and rosiglitazone could inhibit the production of IL-6 and TNF-α induced by latanoprost, respectively. Latanoprost suppressed the expression of differentiation-related mRNA through a positive feedback loop by enhancement of COX-2 expression via FP receptor-activated ERK signaling. The expression of Keratin-17 was upregulated by rosiglitazone and suppressed by LG100268. The application of IL-6 and TNF-α showed negative effects on lipid accumulation in meibomian gland epithelial cells. These results demonstrated that latanoprost could induce inflammation and suppress differentiation of mouse meibomian gland epithelial cells. The activation of PPAR-γ and RXR-α showed an anti-inflammatory effect, showing a potential role to antagonize the effect of latanoprost eyedrops on meibomian gland epithelial cells.

Reverse engineering gene regulatory network based on complex-valued ordinary differential equation model.

BACKGROUND: The growing researches of molecular biology reveal that complex life phenomena have the ability to demonstrating various types of interactions in the level of genomics. To establish the interactions between genes or proteins and understand the intrinsic mechanisms of biological systems have become an urgent need and study hotspot. RESULTS: In order to forecast gene expression data and identify more accurate gene regulatory network, complex-valued version of ordinary differential equation (CVODE) is proposed in this paper. In order to optimize CVODE model, a complex-valued hybrid evolutionary method based on Grammar-guided genetic programming and complex-valued firefly algorithm is presented. CONCLUSIONS: When tested on three real gene expression datasets from E. coli and Human Cell, the experiment results suggest that CVODE model could improve 20-50% prediction accuracy of gene expression data, which could also infer more true-positive regulatory relationships and less false-positive regulations than ordinary differential equation.

NQO2 inhibition relieves reactive oxygen species effects on mouse oocyte meiotic maturation and embryo development.

NRH: quinone oxidoreductase 2 (NQO2) is a cytosolic and ubiquitously expressed flavoprotein that catalyzes the two-electron reduction of quinone to hydroquinones. Herein, we assessed the protein expression, subcellular localization, and possible functions of NQO2 in mouse oocyte meiotic maturation and embryo development. Western blot analysis detected high and stable protein expression of NQO2 in mouse oocytes during meiotic progression. Immunofluorescence illustrated NQO2 distribution on nuclear membrane, chromosomes, and meiotic spindles. Microtubule poisons treatment (nocodazole and taxol) showed that filamentous assembly of NQO2 and its co-localization with microtubules require microtubule integrity and normal dynamics. Increased levels of NQO2, reactive oxygen species (ROS), malondialdehyde (MDA), and autophagy protein Beclin1 expression were detected in oocytes cultured with ROS stimulator vitamin K3 (VK3), combined with decreased antioxidant glutathione (GSH). These oocytes were arrested at metaphase I with abnormal spindle structure and chromosome configuration. However, this impact was counteracted by melatonin or NQO2 inhibitor S29434, and the spindle configuration and first polar body extrusion were restored. Similarly, morpholino oligo-induced NQO2 knockdown suppressed ROS, MDA, and Beclin1, instead increased GSH in oocytes under VK3. Supplementary S29434 or melatonin limited changes in NQO2, ROS, MDA, Beclin1, and GSH during in vitro aging of ovulated oocytes, thereby maintaining spindle structure, as well as ordered chromosome separation and embryo development potential after parthenogenetic activation with SrCl2. Taken together, NQO2 is involved in ROS generation and subsequent cytotoxicity in oocytes, and its inhibition can restore oocyte maturation and embryo development, suggesting NQO2 as a pharmacological target for infertility cure.

Ste20-like kinase activity promotes meiotic resumption and spindle microtubule stability in mouse oocytes.

Ste20-like kinase (SLK) is involved in cell proliferation and migration in somatic cells. This study aims to explore SLK expression and function in mouse oocyte meiosis. Western blot, immunofluorescence, Co-immunoprecipitation, drug treatment, cRNA construct and in vitro transcription, microinjection of morpholino oilgo (MO) and cRNA were performed in oocytes. High and stable protein expression of SLK was detected in mouse oocyte meiosis, with dynamic distribution in the nucleus, chromosomes and spindle apparatus. SLK phosphorylation emerges around meiotic resumption and reaches a peak during metaphase I (MI) and metaphase II. SLK knockdown with MO or expression of kinase-dead SLK K63R dramatically delays meiotic resumption due to sequentially suppressed phosphorylation of Polo-like kinase 1 (Plk1) and cell division cycle 25C (CDC25C) and dephosphorylation of cyclin-dependent kinase 1 (CDK1). SLK depletion promotes ubiquitination-mediated degradation of paxillin, an antagonist to α-tubulin deacetylation, and thus destroys spindle assembly and chromosome alignment; these phenotypes can be substantially rescued by exogenous expression of SLK kinase active fragment. Additionally, exogenous SLK effectively promotes meiotic progression and spindle assembly in aging oocytes with reduced SLK. Collectively, this study reveals SLK is required for meiotic resumption and spindle assembly in mouse oocyte meiosis.

Adverse effects of 2-Methoxyestradiol on mouse oocytes during reproductive aging.

2-Methoxyestradiol (2-ME2) is a metabolite of 17ß-estradiol and is currently in clinical trials as an antitumor agent. Here we found 2-ME2 level remains stable in the local environment of ovaries but declines in serum in aging mice, and exogenous 2-ME2 impacts the meiotic maturation of mouse oocytes in dose-dependent manner. In vitro 2-ME2 application arrested oocytes at metaphase I (MI), with abnormal spindle structure and chromosome alignment. 2-ME2 exposure induced excessive production of reactive oxygen species (ROS) and malondialdehyde, as well as accelerated apoptosis progression. 2-ME2 unbalanced mitochondrial dynamics by increasing DRP1 and MFN1 while decreasing Opa1. Similar phenotypes were also observed in oocytes from mice injected intraperitoneally with 2-ME2. Taken together, this study indicates 2-ME2 exposure impairs oocyte meiotic maturation through inducing mitochondrial imbalance, oxidative stress and apoptosis. The gradual decline in oocyte quality and quantity may be associated with the stable 2-ME2 in ovaries during female reproductive aging.

Stable expression and multi-site location of Odf2 in mouse oocytes, sperm and early embryos.

In mammals, centriole is degenerated during early oogenesis, but it is still not known about the expression and function of centriolar structural components in oocyte meiosis. Here we found that Odf2 (outer dense fiber of sperm tails 2), a key centriolar appendage protein, was stably expressed in mouse oocytes during meiotic progression. Distinct from its single location at centrosomes in somatic mitosis, Odf2 was multiply located at microtubule organizing centers (MTOCs), chromosome centromeres and vesicles in oocyte meiosis. In addition, the vesicle-associated Odf2 disappeared in oocytes treated with the vesicle inhibitor Brefeldin A. Odf2 was mainly co-localized with the mitochondrial sheath in the sperm tail and presented as double spots, similar to γ-tubulin, in the sperm neck region. After fertilization, Odf2 remained on vesicles in embryos from 1-cell to 4-cell stage but was only detected on centrosomes at blastocyst stage. Taken together, Odf2 is expressed precisely in mouse oocytes even in the absence of intact centriole structure, and may regulate oocyte spindle assembly and positioning, additionally, the sperm motility and early embryo development.

Cyclosporine A (0.05%) Ophthalmic Gel in the Treatment of Dry Eye Disease: A Multicenter, Randomized, Double-Masked, Phase III, COSMO Trial.

Purpose: To confirm the efficacy and safety of a novel ophthalmic cyclosporine A gel (CyclAGel, 0.05% CsA) in treating patients with moderate-to-severe dry eye disease (DED). Patients and Methods: The COSMO trial was a randomized, multicenter, double-masked, vehicle-controlled, phase III trial. Patients with moderate-to-severe DED were enrolled in 37 hospitals in China between November 2020 and April 2021. Eligible patients were randomized 1:1 to receive CyclAGel 0.05% or vehicle eye drops once nightly (QD). The primary endpoint was the proportion of subjects with at least a 1-point improvement in ICSS at day 84. Treatment-emergent adverse events (TEAEs) were recorded. Results: The full analysis set (FAS) included 315 and 312 participants in the CyclAGel and vehicle groups, respectively. The primary efficacy endpoint was achieved. The proportion of subjects with at least a 1-point improvement in ICSS from baseline to day 84 was significantly higher in the CyclAGel group than in the vehicle group (73.7% [232/315] vs 53.2% [166/312], P<0.0001). Significant improvements relative to the vehicle were also observed in the ICSS and Oxford scale scoring of corneal and conjunctival fluorescein staining at day 14, 42, and 84. The Schirmer tear test results were significantly higher in the CyclAGel group than in the vehicle group on days 14 and 84 (all P<0.05). The CyclAGel 0.05% was well tolerated, and the TEAEs were mostly mild. The most frequent treatment-related TEAE was eye pain (6.9% vs 1.6% in the CyclAGel and vehicle groups, respectively). No serious treatment-related TEAEs were reported. Conclusion: Clinically and statistically significant improvements in ICSS, tear production, and symptoms were observed in participants administered CyclAGel 0.05% QD for moderate-to-severe DED. CyclAGel 0.05% QD is a new effective, safe, and well-tolerated therapeutic option that might bring additional benefits of convenience and compliance as a once-A-day treatment for DED.

Chemical shift prediction of RNA imino groups: application toward characterizing RNA excited states.

NH groups in proteins or nucleic acids are the most challenging target for chemical shift prediction. Here we show that the RNA base pair triplet motif dictates imino chemical shifts in its central base pair. A lookup table is established that links each type of base pair triplet to experimental chemical shifts of the central base pair, and can be used to predict imino chemical shifts of RNAs to remarkable accuracy. Strikingly, the semiempirical method can well interpret the variations of chemical shifts for different base pair triplets, and is even applicable to non-canonical motifs. This finding opens an avenue for predicting chemical shifts of more complicated RNA motifs. Furthermore, we combine the imino chemical shift prediction with NMR relaxation dispersion experiments targeting both 15N and 1HN of the imino group, and verify a previously characterized excited state of P5abc subdomain including an earlier speculated non-native G•G mismatch.

Efficacy of Immunosuppressants in High Rejection Risk Keratoplasty: A Meta-Analysis of Comparative Studies.

PURPOSE: To evaluate the prophylactic effects of immunosuppressants in corneal graft rejection after high-risk penetrating keratoplasty. METHODS: We searched PubMed, Embase, and the Cochrane Library for comparative studies published between 1989 and 2019 that evaluated the efficacy of immunosuppressants for high-risk corneal graft. The primary outcomes were the 1- and 3-year rejection rates. A fixed-effects or random-effects model was used on the basis of the I2 value, and the results were reported as odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Topical tacrolimus (FK506) was more effective than topical cyclosporine A (CsA) at reducing the 1-year graft rejection rate (OR: 0.17; 95% CI, 0.08-0.37, P<0.01). However, the combination of steroid with either topical FK506 (OR: 0.4; 95% CI, 0.16-1.04, P = 0.09) or CsA (OR: 0.74; 95% CI, 0.32-1.71, P= 0.48) did not show significant superiority in preventing immune rejection compared with steroid monotherapy. Mycophenolate mofetil (MMF) was more effective than CsA at reducing the 1-year graft rejection rate (OR: 2.67; 95% CI, 1.50-4.76, P<0.01). However, MMF was not significantly superior to CsA at reducing the 3-year graft rejection rate (OR: 1.21; 95% CI, 0.45-3.25, P = 0.71). For reducing the 1-year rejection rate, MMF (OR: 0.12; 95% CI, 0.03-0.39, P < 0.01) and CsA (OR: 0.28; 95% CI, 0.10-0.76, P = 0.01) were each more effective than the control groups. CONCLUSIONS: FK506 eye drops, MMF, and systemic CsA were considered to be promising management to prevent rejection in high-risk penetrating keratoplasty in the present study.

Promoter Methylation Regulates ApoA-I Gene Transcription in Chicken Abdominal Adipose Tissue.

As a central constituent of HDL (high-density lipoprotein), apolipoprotein A-I (ApoA-I) has a vital function in lipid metabolism. Our previous studies confirmed that ApoA-I was differentially expressed in the adipose tissue of the abdomen of lean and fat broilers. The aim of the current work was to evaluate whether the transcription of ApoA-I in chicken abdominal adipose tissue was regulated by DNA methylation. The methylation status of ApoA-I promoter CpG island (PCGI) and promoter non-CpG island (PNCGI) as well as the ApoA-I expression level in adipose tissue of lean and fat broilers were determined using Sequenom MassARRAY and real-time PCR. The correlation analysis results showed that the methylation level of PCGI and the ApoA-I mRNA expression level were negatively correlated. Bisulfite sequencing PCR was used to assess the methylation level of ApoA-I promoter in the ICP1 cells treated with 5-aza-2'-deoxycytidine (5-Aza-CdR: an inhibitor of DNA methyltransferase). The result showed that 5-Aza-CdR caused a reduction in the methylation level of the ApoA-I promoter, thereby causing an increase in expression of the ApoA-I mRNA. Meanwhile, luciferase reporter assays indicated that in vitro methylation of the ApoA-I promoter containing CpG island with CpG methyltransferase led to transcriptional repression. Furthermore, the noticeable activation of NRF1 on ApoA-I transcription was largely enhanced  by the demethylation of the ApoA-I PCGI region. These observations indicated that the differential expression of ApoA-I gene in the adipose tissue of broilers could be mediated by transcription regulation, at least in part by DNA methylation in its PCGI region.

PLD2 regulates microtubule stability and spindle migration in mouse oocytes during meiotic division.

Phospholipase D2 (PLD2) is involved in cytoskeletal reorganization, cell migration, cell cycle progression, transcriptional control and vesicle trafficking. There is no evidence about PLD2 function in oocytes during meiosis. Herein, we analyzed PLD2 expression and its relationship with spindle formation and positioning in mouse oocyte meiosis. High protein level of PLD2 was revealed in oocytes by Western blot, which remained consistently stable from prophase I with intact germinal vesicle (GV) up to metaphase II (MII) stage. Immunofluorescence showed that PLD2 appeared and gathered around the condensed chromosomesafter germinal vesicle breakdown (GVBD), and co-localized with spindle from pro-metaphase I (pro-MI) to metaphase I (MI) and at MII stage. During anaphase I (Ana I) to telophase I (Tel I) transition, PLD2 was concentrated in the spindle polar area but absent from the midbody. In oocytes incubated with NFOT, an allosteric and catalytic inhibitor to PLD2, the spindle was enlarged and center-positioned, microtubules were resistant to cold-induced depolymerization and, additionally, the meiotic progression was arrested at MI stage. However, spindle migration could not be totally prevented by PLD2 catalytic specific inhibitors, FIPI and 1-butanol, implying at least partially, that PLD2 effect on spindle migration needs non-catalytic domain participation. NFOT-induced defects also resulted in actin-related molecules' distribution alteration, such as RhoA, phosphatidylinosital 4, 5- biphosphate (PIP2), phosphorylated Colifin and, consequently, unordered F-actin dynamics. Taken together, these data indicate PLD2 is required for the regulation of microtubule dynamics and spindle migration toward the cortex in mammalian oocytes during meiotic progression.

[Clinical analysis of immediate implant and implant-supported complete denture for 12 patients with severe periodontitis].

OBJECTIVE: To evaluate clinical results of implant-supported complete denture of 12 patients with severe chronic periodontitis. METHODS: After systematically periodontic treatment and controlling the inflammation, 12 cases with severe chronic periodontitis were extracted the residual teeth and received immediate implant placement. The implant-supported complete dentures were finished at 5 or 6 months after operation. Restorative results were evaluated with clinical examination, radiological examination and chief complaints of the patients. RESULTS: A total of 108 implants were placed into 20 dental arches, including 37 immediate implants. Average loading was for 3 years and all implants were stable. Progressive bone resorption was observed around two implants. The average bone resorption of other peri-implants was (1.33 +/- 0.10) mm. The implant survival rate was 98.1%. The immediate implant survival rate was 97.3%. CONCLUSIONS: After periodontic treatment, the patients with severe chronic periodontitis could be restored using immediate implant placement and implant-supported complete denture. It can shorten the treatment period, reduce absorption of alveolar and obtain a favorable result by oral health care.

[Clinical study on the stability of palatal implant anchorage].

OBJECTIVE: To evaluate the clinical stability of the palatal implant anchorage system in orthodontic treatment. METHODS: Sixteen osseointegrated implants (5.0 mm in diameter, 6 mm length) were inserted in the median palatal suture area of 19 patients with malocclusion (average age: 18.22 +/- 7.10 years, from 11 years to 35 years 1 month) as anchorage of active orthodontic treatment with MBT appliance. The standard lateral cephalogram after implant placement and before implant removing was taken to compare the radiological parameters. RESULTS: The successful rate of palatal implant anchorage was 84.2%. The average duration of 16 palatal implant was 23.08 +/- 8.06 months (from 10 months to 36 months). There were no statistical differences in all parameters from the implant placement to the end of treatment. IL-X was from (62.88 +/- 5.85) mm to (62.45 +/- 6.70) mm, IL-Y was from (36.66 +/- 5.41) mm to (37.96 +/- 4.90) mm, IAP-PP was from (73.81 +/- 8.84) degrees to (74.72 +/- 9.22) degrees, IAP-Y was from (62.09 +/- 9.33) degrees to (63.85 +/- 10.96) degrees, U6-Y is from (20.80 +/- 5.87) mm to (21.49 +/- 6.00) mm. CONCLUSIONS: Stable palatal implant anchorage was maintained during active orthodontic treatment.