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The progressive degradation in the trabecular meshwork (TM) is related to age-related ocular diseases like primary open-angle glaucoma. However, the molecular basis and biological significance of the aging process in TM have not been fully elucidated. Here, we established a dynamic single-cell transcriptomic landscape of aged macaque TM, wherein we classified the outflow tissue into 12 cell subtypes and identified mitochondrial dysfunction as a prominent feature of TM aging. Furthermore, we divided TM cells into 13 clusters and performed an in-depth analysis on cluster 0, which had the highest aging score and the most significant changes in cell proportions between the two groups. Ultimately, we found that the APOE gene was an important differentially expressed gene in cluster 0 during the aging process, highlighting the close relationship between cell migration and extracellular matrix regulation, and TM function. Our work further demonstrated that silencing the APOE gene could increase migration and reduce apoptosis by releasing the inhibition on the PI3K-AKT pathway and downregulating the expression of extracellular matrix components, thereby increasing the aqueous outflow rate and maintaining intraocular pressure within the normal range. Our work provides valuable insights for future clinical diagnosis and treatment of glaucoma.

Adhesives and glass are commonly used in electronic packaging. The macroscopic and microscopic (molecular-level) structures at the interface between glass and epoxy adhesives have attracted a great deal of interest in the field of surface analysis. In many previous studies, sum frequency generation vibration spectroscopy (SFG-VS) with a total internal reflection (TIR) silica prism was used to probe these types of interfaces. However, for adhesives with relatively high refractive indexes, the requirement of a TIR geometry is not matched, and the origin of SFG-VS signals from the interface involving the prism and adhesive is complicated. Many efforts have been made to resolve this issue. This study provides an approach to analyze the linear and nonlinear optical transmission and emission in a silica–epoxy adhesive–silica sandwich sample with different adhesive thicknesses (i.e., 1.5, 4.5, and 13.5 μm). The origin of SFG-VS signals and structural information in the adhesive were revealed from spectroscopy, morphological, and adhesion tests. It was observed that an insufficient adhesive thickness leads to void structures in the adhesive layer, generating a relatively high SFG-VS signal at the adhesive–air interface. However, samples with a filled adhesive layer emit much weaker SFG signals. Lap shear tests, energy-dispersive X-ray imaging, scanning electron microscopy, and SFG-VS with the introduction of liquids into the sample were used to confirm conclusions obtained from linear and nonlinear optical measurements. This study provides a straightforward experimental approach to analyze the relatively complicated interfaces involving polymers.

ABSTRACTThe objective of this study was to evaluate the shade tolerance of one annual and five perennial Medicagogenotypes under different artificial shade levels and determine the shade tolerant candidates for an intercropping system. The results showed that shade significantly affected branch number and plant dry weight, but not plant height and leaf area. The mean branch number and dry weight plant−1(g) across the tested genotypes were 42.5 and 2.2 under non-shade condition, decreased to 39.5 and 2.0 at 25% shade level, and were further reduced to 17.2 and 0.4 at 75% shade level, respectively. The mean chlorophyll content (mg g−1) and maximum quantum yield of Photosystem II (Fv/Fm) across the tested genotypes at 75% shade level (38.8; 0.6) were significantly lower than other shade levels (46.6–47.3; 0.69–0.70). Antioxidant enzyme activity and malondialdehyde production in Medicagoleaves increased concomitantly with increased shade levels compared to non-shade control. The evaluation showed that annual M. polymorpha‘Jinhuacai’ (range of mean: 0.97–5.97 g) significantly out-yielded those perennials (range of mean: 0.26–2.57 g) by a wide margin at all shade levels. This study would provide baseline information on shade tolerance of Medicagospecies for the selection of intercropped candidates in an intercropping system.

The axons of retinal ganglion cells (RGCs) form the optic nerve, transmitting visual information from the eye to the brain. Damage or loss of RGCs and their axons is the leading cause of visual functional defects in traumatic injury and degenerative diseases such as glaucoma. However, there are no effective clinical treatments for nerve damage in these neurodegenerative diseases. Here, we report that LIM homeodomain transcription factor Lhx2promotes RGC survival and axon regeneration in multiple animal models mimicking glaucoma disease. Furthermore, following N-methyl-D-aspartate (NMDA)-induced excitotoxicity damage of RGCs, Lhx2mitigates the loss of visual signal transduction. Mechanistic analysis revealed that overexpression of Lhx2supports axon regeneration by systematically regulating the transcription of regeneration-related genes and inhibiting transcription of Semaphorin 3C (Sema3C). Collectively, our studies identify a critical role of Lhx2in promoting RGC survival and axon regeneration, providing a promising neural repair strategy for glaucomatous neurodegeneration.

Tb3+,Dy3+-co-doped Ca9TbxDy1–x(PO4)5(SiO4)F2phosphors were prepared via high-temperature solid-phase reaction method and the potential application in optical temperature measurements due to their color-tunable property was investigated in detail. The photoluminescence emission (PL) and photoluminescence excitation (PLE) spectra results show that the as-prepared phosphors exhibit both Tb3+and Dy3+emissions at 546 nm (5D4–7F5transition of Tb3+) and 587 nm (4F9/2–6H13/2transition of Dy3+) upon 376 nm excitation, respectively. In addition, the fluorescence decay analysis shows that the lifetime of the Tb3+emission rapidly decreases, which confirms the energy transfer existence between Dy3+and Tb3+. Under 376 nm excitation, the temperature dependence of the fluorescence intensity ratios for the dual-mission bands peaked at 546 and 587 nm was studied in the temperature range from 303 to 573 K. The results show that with the increase of Dy3+concentration, the relative sensitivity first increases and then decreases, what's more, the maximum relative sensitivity is 3.142 × 10−3%/K for Ca9TbxDy1–x(PO4)5(SiO4)-F2(x = 0.4). As a consequence, this preliminary study provides a novel method for exploring the novel thermometers.

Serine/arginine-protein kinase 1 (SRPK1) regulates alternative splicing of VEGF-A to pro-angiogenic isoforms and SRPK1 inhibition can restore the balance of pro/antiangiogenic isoforms to normal physiological levels. The lack of potency and selectivity of available compounds has limited development of SRPK1 inhibitors, with the control of alternative splicing by splicing factor-specific kinases yet to be translated. We present here compounds that occupy a binding pocket created by the unique helical insert of SRPK1, and trigger a backbone flip in the hinge region, that results in potent (<10 nM) and selective inhibition of SRPK1 kinase activity. Treatment with these inhibitors inhibited SRPK1 activity and phosphorylation of serine/arginine splicing factor 1 (SRSF1), resulting in alternative splicing of VEGF-A from pro-angiogenic to antiangiogenic isoforms. This property resulted in potent inhibition of blood vessel growth in models of choroidal angiogenesis in vivo. This work identifies tool compounds for splice isoform selective targeting of pro-angiogenic VEGF, which may lead to new therapeutic strategies for a diversity of diseases where dysfunctional splicing drives disease development.

The pathogenesis of glaucoma is still unknown. There are few studies on the dynamic change of tissue-specific and time-specific molecular pathophysiology caused by ocular hypertension (OHT). This study aimed to identify the early proteomic alterations in the retina, optic nerve head (ONH), and optic nerve (ON). After establishing a rat model of OHT, we harvested the tissues from control and glaucomatous eyes and analyzed the changes in protein expression using a multiplexed quantitative proteomics approach (TMT-MS3). Our study identified 6403 proteins after 1-day OHT and 4399 proteins after 7-days OHT in the retina, 5493 proteins after 1-day OHT and 4544 proteins after 7-days OHT in ONH, and 5455 proteins after 1-day OHT and 3835 proteins after 7-days OHT in the ON. Of these, 560 and 489 differential proteins were identified on days 1 and 7 after OHT in the retina, 428 and 761 differential proteins were identified on days 1 and 7 after OHT in the ONH, and 257 and 205 differential proteins on days 1 and 7 after OHT in the ON. Computational analysis on days 1 and 7 of OHT revealed that alpha-2 macroglobulin was upregulated across two time points and three tissues stably. The differentially expressed proteins between days 1 and 7 after OHT in the retina, ONH, and ON were associated with glutathione metabolism, mitochondrial dysfunction/oxidative phosphorylation, oxidative stress, microtubule, and crystallin. And the most significant change in retina are crystallins. We validated this proteomic result with the Western blot of crystallin proteins and found that upregulated on day 1 but recovered on day 7 after OHT, which are promising as therapeutic targets. These findings provide insights into the time- and region-order mechanisms that are specifically affected in the retina, ONH, and ON in response to elevated IOP during the early stages.