Low expression of TGF-ß2 and matrilin2 in human aqueous humour with acute primary angle closure.

Primary angle-closure glaucoma (PACG) is the leading cause of irreversible blindness in the world. Angle closure induced by pupil block and secondary iris synechia is the fundamental pathology of the PACG. The molecular mechanisms of angle closure have not yet been clearly illustrated. This study was designed to investigate the protein difference in the aqueous humour and explore new biomarker of the PACG. Aqueous humour (AH) was collected from patients with acute primary angle closure (APAC) and cataract (n = 10 in APAC group) and patients with cataract only (n = 10 in control group). Samples were pooled and measured using label-free proteome technology. Then, the differentially expressed proteins (DEPs) were verified by ELISA using independent AH samples (n = 20 each group). More than 400 proteins were revealed in both groups through proteomics. Comparing the two groups, there were 91DEPs. These proteins participate in biological activities such as inflammation, fibrosis, nerve growth and degeneration and metabolism. We found that the expression of transforming growth factor-ß2 and matrilin2 was downregulated in the APAC group. The two proteins are related to inflammation and extracellular matrix formation, which might be involved in angle closure. This study characterized DEPs in AH of the APAC and found a downregulated protein matrilin2.

Genetic landscape of human mitochondrial genome using whole-genome sequencing.

Increasing evidences suggest that mitochondrial dysfunction is implicated in diseases and aging, and whole-genome sequencing (WGS) is the most unbiased method in analyzing the mitochondrial genome (mtDNA). However, the genetic landscape of mtDNA in the Chinese population has not been fully examined. Here, we described the genetic landscape of mtDNA using WGS data from Chinese individuals (n = 3241). We identified 3892 mtDNA variants, of which 3349 (86%) were rare variants. Interestingly, we observed a trend toward extreme heterogeneity of mtDNA variants. Our study observed a distinct purifying selection on mtDNA, which inhibits the accumulation of harmful heteroplasmies at the individual level: (1) mitochondrial dN/dS ratios were much <1; (2) the dN/dS ratio of heteroplasmies was higher than homoplasmies; (3) heteroplasmies had more indels and predicted deleterious variants than homoplasmies. Furthermore, we found that haplogroup M (20.27%) and D (20.15%) had the highest frequencies in the Chinese population, followed by B (18.51%) and F (16.45%). The number of variants per individual differed across haplogroup groups, with a higher number of homoplasmies for the M lineage. Meanwhile, mtDNA copy number was negatively correlated with age but positively correlated with the female sex. Finally, we developed an mtDNA variation database of Chinese populations called MTCards (http://genemed.tech/mtcards/) to facilitate the query of mtDNA variants in this study. In summary, these findings contribute to different aspects of understanding mtDNA, providing a better understanding of the genetic basis of mitochondrial-related diseases.

Proteomic characteristics of the aqueous humor in Uyghur patients with pseudoexfoliation syndrome and pseudoexfoliative glaucoma.

Pseudoexfoliation syndrome (PEX) is characterized by the deposition of fibrous pseudoexfoliation material (PEXM) in the eye, and secondary glaucoma associated with this syndrome has a faster and more severe clinical course. The incidence of PEX and pseudoexfoliative glaucoma (PEXG) exhibits ethnic clustering; however, few proteomic studies related to PEX and PEXG have been conducted in Asian populations. Therefore, we aimed to conduct proteomic analysis on the aqueous humor (AH) obtained from Uyghur patients with cataracts, those with PEX and cataracts, and those with PEXG and cataracts to better understand the molecular mechanisms of the disease and identify its potential biomarkers. To this end, AH was collected from patients with cataracts (n = 10, control group), PEX with cataracts (n = 10, PEX group), and PEXG with cataracts (n = 10, PEXG group) during phacoemulsification. Label-free quantitative proteomic techniques combined with bioinformatics were used to identify and analyze differentially expressed proteins (DEPs) in the AH of PEX and PEXG groups. Then, independent AH samples (n = 12, each group) were collected to validate DEPs by enzyme-linked immunosorbent assay (ELISA). The PEX group exhibited 25 DEPs, while the PEXG group showed 44 DEPs, both compared to the control group. Subsequently, we found three newly identified proteins in both PEX and PEXG groups, wherein FRAS1-related extracellular matrix protein 2 (FREM2) and osteoclast-associated receptor (OSCAR) exhibited downregulation, whereas coagulation Factor IX (F9) displayed upregulation. Bioinformatics analysis suggested that extracellular matrix interactions, abnormal blood-derived proteins, and lysosomes were mainly involved in the process of PEX and PEXG, and the PPI network further revealed F9 may serve as a potential biomarker for both PEX and PEXG. In conclusion, this study provides new information for understanding the proteomics of AH in PEX and PEXG.

SB431542 partially inhibits high glucose-induced EMT by restoring mitochondrial homeostasis in RPE cells.

BACKGROUND: The epithelial-mesenchymal transition (EMT) of retinal pigment epithelial (RPE) cells participated in the development of retinal fibrosis. SB431542 is a small molecule inhibitor with inhibitory effects on the ALK4, ALK5 and ALK7. Our study aimed to explore the effect of SB431542 on the EMT of RPE cells and to provide new ideas for the treatment of retinal fibrosis. METHODS: We performed fundus fluorescein angiography, optical coherence tomography and hematoxylin-eosin staining in vivo to observe the effect of SB431542 on choroidal neovascularization (CNV)-induced retinopathy. The proliferation, migration, cytoskeleton, adhesion, reactive oxygen species (ROS), mitochondrial morphology and membrane potential of RPE cells were observed in vitro through fluorescein diacetate staining, Cell Counting Kit-8 experiment, wound healing assay, phalloidin staining, immunofluorescence, MitoSOX, DCFH-DA, MitoTracker and JC-10 staining. Western blot, reverse transcription quantitative and immunofluorescence were used to detect the expression of EMT-related markers, pERK1/2, pGSK3ß and ß-catenin. RESULTS: SB431542 significantly alleviated retinopathy in the CNV model. The proliferation, migration and adhesion in RPE cells decreased to a certain extent in SB431542 treatment. SB431542 partially normalized the structure of RPE cells. The expression levels of E-cadherin increased, while the expression levels of laminin and N-cadherin decreased with SB431542 treatment. SB431542 reduced the production of total ROS, mitochondrial SOX and recovered the mitochondrial membrane potential to a certain degree. In addition, our study showed that SB431542 downregulated the phosphorylation of ERK1/2, GSK3ß and the expression of ß-catenin. CONCLUSION: SB431542 improved EMT in RPE cells by maintaining mitochondrial homeostasis via the ERK1/2 and GSK3ß/ß-catenin pathways. Video Abstract SB431542 inhibits EMT in RPE cells under high glucose conditions.

Performance evaluation of differential splicing analysis methods and splicing analytics platform construction.

A proportion of previously defined benign variants or variants of uncertain significance in humans, which are challenging to identify, may induce an abnormal splicing process. An increasing number of methods have been developed to predict splicing variants, but their performance has not been completely evaluated using independent benchmarks. Here, we manually sourced ∼50 000 positive/negative splicing variants from > 8000 studies and selected the independent splicing variants to evaluate the performance of prediction methods. These methods showed different performances in recognizing splicing variants in donor and acceptor regions, reminiscent of different weight coefficient applications to predict novel splicing variants. Of these methods, 66.67% exhibited higher specificities than sensitivities, suggesting that more moderate cut-off values are necessary to distinguish splicing variants. Moreover, the high correlation and consistent prediction ratio validated the feasibility of integration of the splicing prediction method in identifying splicing variants. We developed a splicing analytics platform called SPCards, which curates splicing variants from publications and predicts splicing scores of variants in genomes. SPCards also offers variant-level and gene-level annotation information, including allele frequency, non-synonymous prediction and comprehensive functional information. SPCards is suitable for high-throughput genetic identification of splicing variants, particularly those located in non-canonical splicing regions.

MiR-29a-3p inhibits high-grade transformation and epithelial-mesenchymal transition of lacrimal gland adenoid cystic carcinoma by targeting Quaking.

BACKGROUND: Lacrimal adenoid cystic carcinoma (LACC) is the most common orbital malignant epithelial neoplasm. LACC with high-grade transformation (LACC-HGT) has higher rates of recurrence, metastasis, and mortality than LACC without HGT. This study investigated the effects of microRNA-29a-3p (miR-29a-3p) in the pathogenesis of LACC-HGT. METHODS: An Agilent human miRNA microarray was used to screen the differentially expressed miRNAs (DEMs) in LACC and LACC-HGT tumor tissues. Then, the primary cells obtained in previous studies were used to determine the effect of miR-29a-3p. RESULTS: The expression of miR-29a-3p was abnormally lower in LACC-HGT than in LACC. miR-29a-3p can specifically target the 3' UTR of Quaking mRNA and down-regulate Quaking expression, thereby inhibiting the proliferation, migration, and epithelial-mesenchymal transition of LACC cells. CONCLUSIONS: This study illustrated that miR-29a-3p functions as a tumor suppressor by down-regulating the expression of Quaking to inhibit the tumorigenesis of LACC cells. This study may also reveal the pathogenesis of HGT in LACC cells and provide a reference for LACC-HGT targeted diagnosis.

Coating zirconium oxide-nanocomposite with humic acid for recovery of mercury and chromium in hazardous waste of chemical oxygen demand test.

Hazardous waste of chemical oxygen demand (COD) test (HWCOD) is one of the most common laboratory wastewaters, containing large amounts of H2SO4 and highly toxic Cr3+ and Hg2+. Current treatment methods suffered from incomplete removal of Cr3+ and high-cost. Herein, a humic acid-coated zirconium oxide-resin nanocomposite (HA-HZO-201) was fabricated for efficient recovery of Cr3+ and Hg2+ in HWCOD. The synthesized HA-HZO-201 shows excellent tolerance to wide pH range (1-5) and high salinity (3.5 mol/L NaCl), as well as adsorption capacity for Cr3+ (37.5 mg/g) and Hg2+ (121.3 mg/g). After treating with HA-HZO-201 by using a fixed-bed adsorption procedure, the final Cr3+ and Hg2+ concentrations in HWCOD decreased to 0.28 and 0.02 mg/L, respectively. In addition, the HA-HZO-201 can be regenerated by desorption and recovery of Cr3+ and Hg2+ using HNO3 and thiourea as eluents, respectively. After 5 cycles of adsorption/desorption, the removal efficiencies still reach up to 86.0% for Cr3+ and 89.7% for Hg2+, indicating an excellent regeneration of HA-HZO-201. We hope this work open new opportunities for treatment of HWCOD with high-efficiency and low-cost.

Total Organic Carbon as a Quantitative Index of Micro- and Nano-Plastic Pollution.

The global pollution of micro- and nano-plastics (MNPs) calls for monitoring methods. As diverse mixtures of various sizes, morphologies, and chemical compositions in the environment, MNPs are currently quantified based on mass or number concentrations. Here, we show total organic carbon (TOC) as an index for quantifying the pollution of total MNPs in environmental waters. Two parallel water samples are respectively filtered with a carbon-free glass fiber membrane. Then, one membrane with the collected particulate substances is treated by potassium peroxodisulfate oxidation and Fenton digestion in sequence for quantifying the sum of MNPs and particulate black carbon (PBC) as TOCMNP&PBC using a TOC analyzer, another membrane is treated by sulfonation and Fenton digestion for quantifying PBC as TOCPBC, and the TOC of MNPs is calculated by subtracting TOCPBC from TOCMNP&PBC. The feasibility of our method is demonstrated by determination of various MNPs of representative plastic types and sizes (0.5-100 µm) in tap, river, and sea water samples, with low detection limits (∼7 µg C L-1) and high spiked recoveries (83.7-114%). TOC is a powerful index for routine monitoring of MNP pollution.

Impact of persistent subclinical hypothyroidism on clinical outcomes in non-ST-segment elevation acute coronary syndrome undergoing percutaneous coronary intervention.

BACKGROUND: Data on the association of subclinical hypothyroidism (SCH) with the severity of coronary artery disease and major adverse cardiovascular and cerebral events (MACCE) in patients with non-ST-segment elevation acute coronary syndrome (NSTE-ACS) after percutaneous coronary intervention (PCI) are limited and conflicting. OBJECTIVE: We established the baseline rate of SCH and followed the trajectory of thyroid-stimulating hormone (TSH) values during and after hospitalisation for PCI for up to six months and determined whether persistent SCH was associated with the severity of coronary artery disease and MACCE in patients with NSTE-ACS after PCI. DESIGN: Population-based prospective cohort study. PATIENTS: We included patients with NSTE-ACS who underwent PCI with simple balloon angioplasty or stent implantation for coronary heart disease. MEASUREMENTS: Thyroid function tests of patients before PCI and 1 day, 1 week, 1 and 6 months after PCI were performed. Cases showing transient SCH were excluded. Patients were divided into two groups based on the results of four TSH tests: 0.27-4.2 mIU/L (n = 1472, 89.7%) and >4.2 mIU/L (n = 170, 10.4%). The risk factors for the severity of coronary artery lesions were estimated using multinomial logistic regression analysis. Univariate and multivariate Cox regression analyses were used to study the relationship between TSH and MACCE. RESULTS: Among 1642 patients, there were 1070 males (65.2%) and 572 females (34.8%), with an average age of 62.5 ± 9.6 years. SCH patients had a wider range of diseased vessels and a higher number of diseased vessels (p < .05). TSH level was an independent risk factor for moderate [odds ratio (OR) = 1.144, 95% confidence interval (95% CI): 1.057-1.237, p = .001] and severe (OR = 1.131, 95% CI: 1.043-1.226, p = .003) coronary artery lesions. After adjusting for covariates, the risk of MACCE [hazard ratio (HR): 4.067, p < .001], nonfatal myocardial infarction (HR: 14.724, p = .003), and unplanned PCI (HR: 5.028, p < .001) were higher in the SCH group than in the euthyroidism group. There were no significant differences in the incidence of heart failure (HR: 6.012, p = .175), nonfatal stroke (HR: 2.039, p = .302), unplanned coronary artery bypass grafting (CABG) (HR: 1.541, p = .57), or cardiac death (HR: 2.704, p = .375) between the two groups. CONCLUSIONS: Preoperative TSH levels and changes in thyroid hormone levels several months post-PCI in NSTE-ACS patients are highly significant in practice. Persistent SCH is associated with severe coronary artery lesions and MACCE, and may be a predictor for evaluating the prognosis of PCI-treated NSTE-ACS patients.

AKT2 identified as a potential target of mir-29a-3p via microRNA profiling of patients with high proliferation lacrimal gland adenoid cystic carcinoma.

The lacrimal gland adenoid cystic carcinoma (LACC) is a major orbital malignancy. The recurrence rate and mortality rate are higher in high proliferation LACC(HP-LACC) compared with low proliferation LACC(LP-LACC). In this study, miRNA microarray was used to explore the differentially expressed miRNAs profiling between HP-LACC and LP-LACC and its potential signaling pathway. Tissues from 17 patients with LACC were collected and made into tissue microarrays. Patients were divided into a high proliferation group and a low proliferation group based on Ki-67 value. HE, immunofluorescence (IF), and Immunohistochemistry (IHC) were performed on the tissue microarrays. Eight LACC tissues(4 HP-LACC and 4 LP-LACC) were made into miRNA microarrays and analyzed for miRNA profiles. Differentially expressed miRNAs were analyzed by volcano plot and heat map. Target gene were predicted using the miRWalk and miRDB for these differentially expressed miRNAs, the intersection of the results are used as targets for further gene ontology and KEGG pathway analysis.The four differentially expressed miRNAs were validated by qRT-PCR, the miRNAs with statistically significant differences validated by dual luciferase reporter and qRT-PCR. Finally, IHC was used for their downstream signaling pathway proteins.HE staining showed the presence of tubular, cribriform, and basaloid structures in LACC. IF showed the presence of CK7,P63 fluorescence expression in all three structures.Patients were divided into HP-LACC and LP-LACC based on Ki-67 median value of 11%. IHC and survival analysis showed with the increase of KI-67 ratio, the proportion of P63 decreased, and the expression of P53 increased. The disease-free survival and overall survival of the patients decreased. IHC and survival analysis showed as Ki-67 expression increased, P63 expression decreased, P53 expression elevated, with prognosis worse. Heat map and volcano plot yielded 15 differentially expressed miRNAs between HP-LACC and LP-LACC.The 15 differential miRNAs were used to predict target genes in miRWalk and miRDB databases respectively, and there were 559 target genes after intersection.559 predicted target genes obtained. Go and KEGG analysis showed that these target genes exerted important biological functions through multiple signaling pathways. Among the 15 differentially expressed miRNAs, miR-29a-3p was verified to be significant by qRT-PCR. Dual luciferase reporter and tissue microarray immunohistochemical assays validated that AKT2 was a direct target gene of miR-29a-3p. Current studies have identified differentially expressed miRNAs associated with LACCs of variable proliferation ability, and found that AKT2 is a direct target gene of miR-29a-3p, which will contribute to target gene therapy in patients with high proliferation LACC in the future.

Legumain affects the PI3K/AKT tumor progression pathway in retinoblastoma.

Known as a common malignant tumor among children, retinoblastoma (RB) is highly malignant and has poor prognosis, damages children's vision and degrades quality of life. To identify a potential molecular mechanism of RB, we conducted this study on legumain (LGMN), which is highly expressed in multiple tumors. In this study, we found that LGMN was significantly upregulated in RB cells and was positively expressed in RB tissues. We confirmed that LGMN overexpression (LGMN-OE) can promote RB cell proliferation and inhibit cell apoptosis through CCK8 experiments and flow cytometry. In addition, real-time quantitative polymerase chain reaction (RT‒qPCR) and Western blot results showed that LGMN-OE could regulate the expression of epithelial-mesenchymal transformation-related genes and proteins, related to tumor invasion and metastasis. Moreover, after LGMN knock down, the result was the opposite., RNA sequence analysis revealed 1159 differentially expressed genes between LGMN-OE and the negative control (NCOE), of which 564 were upregulated and 595 were downregulated. The first 10 genes were verified by RT‒qPCR based on P value and fold change. Interestingly, we found that LGMN could regulate the expression of recoverin (RCVRN)through a gene responsible for cancer-related retinopathy. We also screened and verified that LGMN partially activated the PI3K/AKT pathway in RB. Furthermore, we evaluated the effect of legumain inhibitors (e.g., esomeprazole) on RB, and the results suggest that esomeprazole may provide a reference for the clinical adjuvant treatment of RB. In conclusion, legumain can serve as an attractive target for RB therapy and hopefully provide new insights and ideas for the development of targeted drugs and precise personalized clinical therapy.

Low expression of RBP4 in the vitreous humour of patients with proliferative diabetic retinopathy who underwent Conbercept intravitreal injection.

Intravitreal injection of anti-VEGF antibodies has been widely used in the treatment of proliferative diabetic retinopathy (PDR). However, anti-VEGF drugs can exacerbate fibrosis and eventually lead to retinal detachment. To explore proteins closely related to fibrosis, we conducted proteomic analysis of human vitreous humour collected from PDR patients who have or have not intravitreal Conbercept (IVC) injection. Sixteen vitreous humour samples from PDR patients with preoperative IVC and 20 samples from those without preoperative IVC were examined. An immunodepletion kit was used to remove high-abundance vitreous proteins. Conbercept-induced changes were determined using a tandem mass tag-based quantitative proteomic strategy. Enzyme-linked immunosorbent assays were performed to confirm the concentrations of selected proteins and validate the proteomic results. Based on a false discovery rate between 0.05% and -0.05% and a fold-change > 1.5, 97 proteins were altered (49 higher levels and 48 lower levels) in response to IVC. Differentially expressed proteins were found in the extracellular and intracellular regions and were found to be involved in VEGF binding and VEGF-activated receptor activity. Protein-protein interactions indicated associations with fibrosis, neovascularisation and inflammatory signalling pathways. We found the low levels of RBP4 in the vitreous humour of PDR patients with IVC injection, as revealed by ELISA and proteomic profiling. Moreover, RBP4 significantly restored the mitochondrial function of HRMECs induced by AGEs and down regulated the level of glycolysis. Our study is the first to report that RBP4 decreases in the vitreous humour of PDR patients who underwent Conbercept treatment, thereby verifying the role of RBP4 in glucose metabolism. Results provide evidence for the potential mechanism underlying Conbercept-related fibrosis.

Swelling-Induced Fragmentation and Polymer Leakage of Nanoplastics in Seawater.

Nanoplastics (NPs) have been successively detected in different environmental matrixes and have aroused great concern worldwide. However, the fate of NPs in real environments such as seawater remains unclear, impeding their environmental risk assessment. Herein, multiple techniques were employed to monitor the particle number concentration, size, and morphology evolution of polystyrene NPs in seawater under simulated sunlight over a time course of 29 days. Aggregation was found to be a continuous process that occurred constantly and was markedly promoted by light irradiation. Moreover, the occurrence of NP swelling, fragmentation, and polymer leaching was evidenced by both transmission electron microscopy and scanning electron microscopy techniques. The statistical results of different transformation types suggested that swelling induces fragmentation and polymer leakage and that light irradiation plays a positive but not decisive role in this transformation. The observation of fragmentation and polymer leakage of poly(methyl methacrylate) and poly(vinyl chloride) NPs suggests that these transformation processes are general for NPs of different polymer types. Facilitated by the increase of surface functional groups, the ions in seawater could penetrate into NPs and then stretch the polymer structure, leading to the swelling phenomenon and other transformations.

Monitoring the Cd2+ release from Cd-containing quantum dots in simulated body fluids by size exclusion chromatography coupled with ICP-MS.

Quantification of Cd2+ release from Cd-containing quantum dots (QDs) is of fundamental importance to elucidate its toxicity to organisms, but remains a great challenge due to the lack of appropriate analytical method. Herein, a facile method based on size exclusion chromatography (SEC) combined with inductively coupled plasma mass spectrometry (ICP-MS) was developed for separating and quantifying the QDs and counterpart ions. By using the mixture of sodium dodecyl sulfate (SDS) and ethylenediaminetetraacetic acid tetrasodium salt (EDTA) as the mobile phase, the defect of QD and ion adsorption onto the SEC column was overcome, thus realizing the accurate quantification of ionic species. Besides, the concentration of QDs was achieved through subtracting the ion concentration from the total concentration. Selecting CdSe@ZnS as the typical QDs, the Cd2+ release process in four typical simulated body fluids, namely, simulated gastric fluid, simulated sweat, Gamble's solution, and artificial lysosomal fluid, was monitored using the developed SEC-ICP-MS method. The media pH is identified as the decisive factor which controls the dissolution of ZnS shells and also the Cd2+ release kinetics and final concentration. Our results suggest that the oral pathway for QD uptake poses the biggest risk to human health.

Low expression of GSTP1 in the aqueous humour of patients with primary open-angle glaucoma.

Primary open-angle glaucoma (POAG) is characterized by irreversible neurodegeneration accompanied by visual field defects and high intraocular pressure. Currently, an effective treatment is not available to prevent the progression of POAG, other than treatments to decrease the high intraocular pressure. We performed proteomic analysis of aqueous humour (AH) samples from patients with POAG combined with cataract and patients with cataract to obtain a better understanding of the pathogenesis of POAG and explore potential treatment targets for this condition. Samples were collected from 10 patients with POAG combined with cataract and 10 patients with cataract. Samples from each group were pooled. A high-resolution, label-free, liquid chromatography-tandem mass spectrometry-based quantitative proteomic analysis was performed. In total, 610 proteins were identified in human AH samples from the two groups. A total of 48 up-regulated proteins and 49 down-regulated proteins were identified in the POAG combined with cataract group compared with the control group. Gene Ontology (GO) analysis revealed key roles for these proteins in inflammation, immune responses, growth and development, cellular movement and vesicle-mediated transport in the biological process category. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated the down-regulated expression of glutathione S-transferase P (GSTP1) in the glutathione metabolism signalling pathway in the POAG combined with cataract group. Additionally, certain significantly differentially expressed proteins in the proteomic profile were verified by enzyme-linked immunosorbent assay (ELISA). GSTP1 levels were reduced in the human AH samples from the POAG combined with cataract group, based on the results of ELISA and proteomic profiling. Therefore, GSTP1, a redox-related marker, may be involved in the pathological process of POAG and may become a treatment target in the future.

Digestive Elimination of Coexisting Microplastics for Determination of Particulate Black Carbon in Environmental Waters.

Determination of particulate black carbon (PBC) in the environment is of great importance but faces a new challenge due to the increasing occurrence of coexisting microplastics (MPs), which are an emerging contaminant with properties very similar to those of PBC and cannot be discriminated in the chemical digestion procedure of the reported PBC analysis method. Herein, a comprehensive method has been developed for accurately determining PBC by digestive elimination of the coexisting MPs and other non-black carbon organic matter. Water samples were filtered with a glass fiber membrane (0.3 µm pore size), and the collected substances with the membrane were subjected to sulfonation with chlorosulfonic acid and Fenton digestion in sequence and then to the total organic carbon analyzer for quantification of PBC. Under the optimized conditions, MPs of various sizes and polymer types were efficiently eliminated (>91.0%), whereas various PBC samples were undigested with recoveries over 91.7% except for the relatively low recovery of 65.6% for the PBC prepared at a low pyrolysis temperature of 400 °C. The feasibility of the proposed method was verified by analysis of real water samples with a spike recovery of 88.6-100.2%. We anticipate that this work will pave an avenue for reliable determination of PBC in the presence of MPs.

Sequential Isolation of Microplastics and Nanoplastics in Environmental Waters by Membrane Filtration, Followed by Cloud-Point Extraction.

Respective detection of microplastics (MPs) and nanoplastics (NPs) is of great importance for their different environmental behaviors and toxicities. Using spherical polystyrene (PS) and poly(methyl methacrylate) (PMMA) plastics as models, the efficiency for sequential isolation of MPs and NPs by membrane filtration and cloud-point extraction was evaluated. After filtering through a glass membrane (1 µm pore size), over 90.7% of MPs were trapped on the membrane, whereas above 93.0% of NPs remained in the filtrate. The collected MPs together with the glass membrane were frozen in liquid nitrogen, ground, and suspended in water (1 mL) and subjected to pyrolysis-gas chromatography-mass spectrometry (Py-GC/MS) determination. The NPs in the filtrate were concentrated by cloud-point extraction, heated at 190 °C to degrade the extractant, and then determined by Py-GC/MS. For MPs and NPs spiked in pure water, the method detection limits are in the range of 0.05-1.9 µg/L. The proposed method is applied to analyze four real water samples, with the detection of 1.6-7.6 µg/L PS MPs and 0.6 µg/L PMMA MPs in three samples, and spiked recoveries of 75.0-102% for MPs and 67.8-87.2% for NPs. Our method offers a novel sample pretreatment approach for the respective determination of MPs and NPs.

Comparison of A-Scan ultrasonography and the Lenstar optical biometer in Guinea pig eyes.

OBJECTIVE: To compare the biometric parameters provided by A-scan ultrasonography and the Lenstar optical biometer in guinea pig eyes, including anterior segment depth (ASD), lens thickness (LT), vitreous chamber depth (VCD), and axial length (AL), and differences of them between treated form deprivation (FD) eyes and untreated fellow eyes after 4 weeks of FD. METHODS: Three-week-old guinea pigs (N = 41) were subjected to biometric measurements before monocular FD (baseline) and after a 4-week FD. Statistical analyses including within-subject standard deviation (SDwithin), coefficient of variation (CV), and intraclass correlation coefficient (ICC), used to evaluate repeatability for both the A-scan ultrasonography and the Lenstar individually, and correlation and Bland-Altman analyses were used to assess agreement between the two methods. The absolute values of ASD, LT, VCD and AL as measured by the two devices were compared, and the differences of them between treated (T) and untreated fellow (F) eyes (ΔASD, ΔLT, ΔVCD and ΔAL) (Δ = T-F) were compared between the two devices after 4 weeks of FD. RESULTS: Measurements by the Lenstar (ICC: 0.923-0.994) were more repeatable than A-scan ultrasonography (ICC: 0.825-0.870). There was a high correlation for AL (r = 0.851, P < 0.001), a moderate correlation for VCD (r = 0.571, P < 0.001) and LT (r = 0.423, P < 0.001), and a low correlation for ASD (r = 0.230, P < 0.01) between the two devices. The values for ASD, VCD and AL measured by A-scan ultrasonography were larger than those measured by the Lenstar (all, P < 0.001), while LT provided by A-scan ultrasonography was much smaller than that of the Lenstar (P < 0.001). Bland-Altman plots showed poor agreement of absolute values of the four parameters between the two devices. Moreover, there was a high correlation between both methods for ΔAL (r = 0.704, P < 0.001), a moderate correlation for ΔVCD (r = 0.534, P < 0.001) and ΔASD (r = 0.574, P < 0.001), and no correlation for ΔLT (r = 0.303, P = 0.054). The ΔASD, ΔLT, and ΔAL measurements obtained by A-scan ultrasonography were greater than those obtained by the Lenstar (all, P < 0.001), while ΔVCD was mildly smaller using A-scan ultrasonography (P < 0.05). Bland-Altman plots illustrated there is good agreement of ΔAL, ΔVCD, ΔASD, and ΔLT between the two devices. CONCLUSIONS: The Lenstar exhibited better repeatability and provided smaller measurements for AL, VCD and ASD than A-scan ultrasonography. Furthermore, a high correlation and a good agreement for the ΔAL was observed between the two devices after a period of FD. In summary, the two devices cannot replace each other directly to obtain absolute values of ASD, LT, VCD and AL, but the Lenstar still can serve as an option in measuring ΔAL between eyes in guinea pig myopia model.

PSF functions as a repressor of hypoxia-induced angiogenesis by promoting mitochondrial function.

BACKGROUND: Abnormal neovascularization is the most common cause of blindness, and hypoxia alters tissue metabolism, function, and morphology. HIF-1α, the transcriptional activator of VEGF, has intricate mechanisms of nuclear translocation and activation, but its signal termination mechanisms remain unclear. METHODS: We investigated the role of polypyrimidine tract-binding protein-associated splicing factor (PSF) in cellular energy production, migration, and proliferation by targeting HIF-1α in vivo and in vitro PSF plasmids were transfected with liposome 2000 transfection reagent. Young C57/BL6J mice were kept in a hyperoxia environment, followed by indoor air, resulting in oxygen-induced retinopathy. Oxygen-induced retinopathy (OIR) animals were randomly divided into three groups: OIR group, OIR + vector group (OIR cubs treated with rAAV vector) and OIR + PSF group (OIR cubs treated with rAAV-PSF). Age-matched C57/BL6J mice were used as controls and exposed to constant normoxic conditions. The animals were executed and their pupils were subjected to subsequent experiments. The metabolic spectrum was analyzed by Seahorse XFe96 flux analyzer, and OCR and extracellular acidification rate were quantified at the same time. RESULTS: PSF ameliorated retinal neovascularization and corrected abnormal VEGF expression in mice with oxygen-induced retinopathy and reduced intra-retinal neovascularization in Vldlr - / - mice. PSF reprogrammed mitochondrial bioenergetics and inhibited the transition of endothelial cells after hypoxia, suggesting its involvement in pathological angiogenesis.Ectopic PSF expression inhibited hypoxia-induced HIF-1α activation in the nucleus by recruiting Hakai to the PSF/HIF-1α complex, causing HIF-1α inhibition. PSF knockdown increased hypoxia-stimulated HIF-1α reactions. These hypoxia-dependent processes may play a vital role in cell metabolism, migration, and proliferation. Thus, PSF is a potential treatment target in neovascularization-associated ophthalmopathy. CONCLUSION: This is the first study showing that PSF inhibits HIF-1α via recruitment of Hakai, modulates mitochondrial oxidation and glycolysis, and downregulates VEGF expression under hypoxia. We propose a new HIF-1 α/Hakai regulatory mechanism that may play a vital role in the pathogenesis of neovascularization in ophthalmopathy. PSF-Hakai-HIF-1α signaling pathway under hypoxia condition. Schematic diagram showing that the PSF-Hakai-HIF-1α signaling pathway. Under hypoxia condition, PSF-Hakai complex regulate HIF-1α signaling, thus inhibiting downstream target gene VEGF, cell metabolism and angiogenesis eventually. Video Abstract: Detailed information of Materials and Methods.

Highly stable fluorescent probe based on mesoporous silica coated quantum dots for sensitive and selective detection of Cd2.

Cadmium ions have been of crucial concern due to the high biological toxicity and serious environmental risks. Various fluorescent Cd-sensitive probes have been reported with improved sensing properties, but still severely suffer from poor stability and insufficient selectivity. In this work, a stable fluorescent probe based on silica encapsulated quantum dots (QDs) have been developed for rapid, sensitive and selective detection of cadmium ion. To improve fluorescence stability, the strategy of mesoporous silica encapsulation was adopted, in which the mesoporous silica shell offers numerous channels for Cd2+. Further, the Forster Resonance Energy Transfer (FRET) system, where QDs@mSiO2and rhodamine B (RB) are used as donors and acceptors respectively, has been constructed, in which the mesoporous silica shell also serves as spacers with tunable thickness for controlling the QD-RB distance. Under optimal conditions, the probes possess a sensitive fluorescence response with a detection limit of 1.25µM. Visual detection can be realized by the obvious fluorescence changes of the FRET system. In addition, the FRET system shows promising sensing performances both in tap water samples and rice-washed water samples, confirming a great potential for practical application.