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AIM: To analyze the spectrum of isolated pathogens and antibiotic resistance for ocular infections within 5y at two tertiary hospitals in east China. METHODS: Ocular specimen data were collected from January 2019 to October 2023. The pathogen spectrum and positive culture rate for different infection location, such as keratitis, endophthalmitis, and periocular infections, along with antibiotic resistance were analyzed. RESULTS: We included 2727 specimens, including 827 (30.33%) positive cultures. A total of 871 strains were isolated, 530 (60.85%) bacterial and 341 (39.15%) fungal strains were isolated. Gram-positive cocci (GPC) were the most common ocular pathogens. The most common bacterial isolates were Staphylococcus epidermidis (25.03%), Staphylococcus aureus (7.46%), Streptococcus pneumoniae (4.59%), Corynebacterium macginleyi (3.44%), and Pseudomonas aeruginosa (3.33%). The most common fungal genera were Fusarium spp. (12.74%), Aspergillus spp. (6.54%), and Scedosporium spp. (5.74%). Staphylococcus epidermidis strains showed more than 50% resistance to fluoroquinolones. Streptococcus pneumoniae and Corynebacterium macginleyi showed more than 90% resistance to erythromycin. The percentage of bacteria showing multidrug resistance (MDR) significantly decreased (χ2=17.44, P=0.002). CONCLUSION: GPC are the most common ocular pathogens. Corynebacterium macginleyi, as the fourth common bacterium, may currently be the local microbiological feature of east China. Fusarium spp. is the most common fungus. More than 50% of the GPC are resistant to fluoroquinolones, penicillins, and macrolides. However, the proportion of MDR strains has been reduced over time.

AIM: To investigate the effect of age on the expression of Na+-K+-ATPase and acute reversible lens opacification induced by chloral hydrate in mice. METHODS: Acute reversible lens opacification was induced by intraperitoneal injection of 4% chloral hydrate(400mg/kg)in 3-month-old(young group)and 24-month-old(old group)C57BL/6 mice. The lens opacification was graded at 10, 20, 30, 45, 60, 90, 120 and 150min after chloral hydrate injection. The histopathological changes of lens were observed by hematoxylin eosin staining, and the expression of Na+-K+-ATPase in lens was detected by immunohistochemistry. RESULTS: The development of lens opacity is similar in young and old mice after chloral hydrate injection. The lens opacification in the young group appeared earlier, thicker and lasted longer than the old group. HE staining showed that many vesicles appeared in the cortex below lens epithelial cells(LECs), and the structure of superficial lens fiber cells were disordered after chloral hydrate injection. Immunohistochemical staining showed that the expression of Na+-K+-ATPase was positive in LECs and fibers. The expression of Na+-K+-ATPase in LECs were weak before chloral hydrate injection and up-regulated 45min after chloral hydrate injection in young and old groups. The up-regulation of Na+-K+-ATPase was stronger in the old group than in the young group. CONCLUSION: Age may play a role in the acute reversible lens opacification induced by chloral hydrate in mice. The expression of Na+-K+-ATPase is involved in lens opacity induced by chloral hydrate.

AIM: To investigate the effects of doxycycline(DOX)on vasculogenic mimicry(VM)in human pterygium fibroblasts(HPFs)and its molecular mechanisms.METHODS: Primary cultured HPFs were identified by Vimentin and CK through immunocytochemical staining. HPFs were divided into control group and DOX group including low, medium and high concentrations(50, 100, 200mg/L). The activity and migration of HPFs were detected by cell counting kit-8(CCK-8)and wound healing assay. The density of VM was observed by three-dimensional cell culture and periodic acid schiff(PAS)staining and compared the differences of VM formation in each group. Western blot was used to analyze the expression of matrix metalloproteinase-9(MMP-9)and vascular endothelial growth factor(VEGF).RESULTS:Immunocytochemical staining results showed that the cells were spindle shaped, meanwhile, they were positive for Vimentin and negative for CK, which were consistent with the characteristics of fibroblasts. Compared with the control group, the cell activity, mobility, VM density and the expression of MMP-9 and VEGF proteins in the DOX group were significantly decreased(P

Objective To investigate the effect of methyltransferase like 3 (METTL3) on the migration of human umbilical vein endothelial cells (HUVECs), and explore the underlying mechanism preliminarily. Methods The sgRNA targeting the first exon of METTL3 was designed with aid of online platform and synthesized, and then inserted into the lentiCRISPR v2 vector. Western blotting and immunofluorescence staining were used to determine the protein expression of METTL3. Cell migration were detected by cell scratch test and transwell chamber assay. Methylated RNA immunoprecipitation was employed to detect m6A modification of cell migration related gene, tyrosine kinase, LYN mRNA. And fluorescent quantitative PCR (qPCR) was adopted to detect the mRNA level of LYN. After recognition proteins IGF2BP1 and IGF2BP2 were knocked down with shRNA technique and YTHDF2 with siRNA, the expression levels of the 3 knockdown molecules and LYN were detected by qPCR. The interaction between LYN mRNA with IGF2BP2 was verified by RNA immunoprecipitation (RIP) assay. qPCR was used to detect the half-life of LYN mRNA after actinomycin D was employed to inhibit the process of transcription. Results The HUVECs with METTL3 stable knockdown were successfully generated by CRISPR-Cas9. METTL3 knockdown resulted in obviously inhibited cell migration (P0.05), while knockdown of IGF2BP2 decreased the LYN mRNA level (P

Purpose. To identify the ability of releasing and uptaking exosomes in rat lens and characterize the exosomal microRNA profile of lens-derived exosomes. Methods. The rat lenses were cultured ex vivo and the medium was collected. The exosomes were isolated from medium and measured in size and concentration by nanoflow cytometry (nFCM) and transmission electron microscopy (TEM) and verified with CD63 and TSG101 by Western blot. The miRNAs in exosomes released from lens epithelial cells (LECs) were sequenced. The plasma exosomes labeled by PKH26 were used to verify the exosomes uptake LECs, and their colocalized fluorescence was imaged by confocal microscopy. Results. LECs released numerous exosomes into the medium through the capsule, which contained abundant miRNAs. The most abundant miRNAs included miR-184, let-7c-5p, let-7a-5p, let-7b-5p, let-7f-5p, miR-125a-5p, miR-204-5p, miR-125b-5p, miR-1b, and miR-23a-3p. The LECs but not the lens fibre cells showed exosome uptake. The LECs uptake more PKH26-labeled exosomes at day 7 than day 3 and day 14. Conclusions. Our results suggested that LECs can release and uptake exosomes through the capsule. Exosomes may be an important way for the lens to communicate among LECs, aqueous humour, vitreous body, and other ocular tissues.

Microplastics (the term for plastics at sizes of p > 0.05) between the sampling station and period for the presence of a microplastics amount. Most of the microplastics were found in fiber form (81.5%), whereas the remaining comprised fragment (18.31%) and film (0.19%) forms. Further analysis with Py-GC/MS under a selective ion monitoring mode indicated that pyrolytic products and fragment ions for a variety of polymers, such as polyvinyl chloride, polypropylene, polyethylene, polyethylene terephthalate, polyamide, and polymethylmethacrylate, were detected. This study provides an insightful application of Py-GC/MS techniques for microplastics monitoring, especially when dealing with analytical amounts of samples.

Age-related cataract (ARC) is a leading cause of visual impairment and blindness worldwide. DNA damage and malfunction of DNA repair are believed to contribute to the pathogenesis of ARC. Aside from increasing age, the risk factors for ARC appear to be rather complex, and one or more gene variations could play critical roles in the diverse processes of ARC progression. This study aimed to investigate the combined effects of different genetic variants on ARC risk.A cohort of 789 ARC patients and 531 normal controls from the Jiangsu Eye Study was included in this study. Genotyping of 18 single-nucleotide polymorphisms (SNPs) in 4 DNA damage/repair genes was performed using TaqMan SNP assays. SNP-SNP interactions were analyzed via multifactor dimensionality reduction (MDR), classification and regression tree (CART) and genetic risk score (GRS) analyses.Based on single-locus analyses of the 18 SNPs examined, WRN-rs11574311 (T>C) was associated with ARC risk. However, in MDR, the gene-gene interaction among the five SNPs (WRN-rs4733220 (G>A), WRN-rs1801195 (T>G), OGG1-rs2072668 (G>C) and OGG1-rs2304277 (A>G)) on ARC risk was significant (OR = 5.03, 95% CI: 3.54~7.13). CART analyses also revealed that the combination of five SNPs above was the best polymorphic signature for discriminating between the cases and the controls. The overall odds ratio for CART ranged from 4.56 to 7.90 showing an incremental risk for ARC. This result indicated that these critical SNPs participate in complex interactions. The GRS results showed an increased risk for ARC among individuals with the SNPs in this polymorphic signature.The use of multifactorial analysis (or an integrated approach) rather than a single methodology could be an improved strategy for identifying complex gene interactions. The multifactorial approach used in this study has the potential to identify complex biological relationships among ARC-related genes and processes. This approach will lead to the discovery of novel biological information, ultimately improving ARC risk management.

Purpose. Lanosterol synthase (LSS) abnormity contributes to lens opacity in rats, mice, dogs, and human congenital cataract development. This study examined whether LSS pathway has a role in different subtypes of age-related cataract (ARC). Methods. A total of 390 patients with ARC and 88 age-matched non-ARC patients were enrolled in this study. LSS expression was analyzed by western blot and enzyme-linked immunosorbent assay (ELISA). To further examine the function of LSS, we used U18666A, an LSS inhibitor in rat lens culture system. Results. In lens epithelial cells (LECs), LSS expression in LECs increased with opaque degree C II, while it decreased with opaque degree C IV and C V. While in the cortex of age-related cortical cataract (ARCC), LSS expression was negatively related to opaque degree, while lanosterol level was positively correlated to opaque degree. No obvious change in both LSS and lanosterol level was found in either LECs or the cortex of age-related nuclear cataract (ARNC) and age-related posterior subcapsular cataract (ARPSC). In vitro, inhibiting LSS activity induced rat lens opacity and lanosterol effectively delayed the occurrence of lens opacity. Conclusions. This study indicated that LSS and lanosterol were localized in the lens of human ARC, including ARCC, ARNC, and ARPSC. LSS and lanosterol level are only correlated with opaque degree of ARCC. Furthermore, activated LSS pathway in lens is protective for lens transparency in cortical cataract.

The formation of primary angle closure (PAC) and primary angle closure glaucoma (PACG) is regulated by a tissue remodeling pathway that plays a critical role in eye development. MicroRNAs (miRNAs) are powerful gene expression regulators and may exert their effects on tissue remodeling genes. This study investigated the associations between gene variants (single-nucleotide polymorphism, SNP) in miRNA binding sites in the 3'-UTR region of genes involved in eye development and PAC.The sample consisted of 232 PAC subjects and 306 controls obtained from a population-based cohort in the Funing District of Jiangsu, China. The markers include 9 SNPs in the COL11A1, PCMTD1, ZNRF3, MTHFR, and ALPPL2 genes respectively. SNP genotyping was performed with a TaqMan-MGB probe using an RT-PCR system.Of the 9 SNPs studied, the frequency of the minor A allele of COL11A1 rs1031820 was higher in the PAC group than in the control group in allele analysis (p = 0.047). The genotype analysis indicated that MTHFR rs1537514 is marginally associated with PAC (p = 0.014). The CC genotype of rs1537514 was present solely in the PAC group. However, the differences lost significance after Bonferroni correction.Our study reveals a possible association of COL11A1 and MTHFR with PAC in the Han Chinese population. These results will contribute to an improved understanding of the genetic basis of PACG.

Chemotherapy resistance is a major obstacle to effective cancer treatment, and promotion of ferroptosis can suppress cisplatin resistance in tumor cells. TCF12 plays a suppressive role in oral squamous cell carcinoma (OSCC), but whether it participates in the regulation of cisplatin resistance by modulating ferroptosis remains unclear. Here, we found that TCF12 expression was decreased in OSCC cells compared with normal oral cells, and it was reduced in cisplatin (DDP)‐resistant OSCC cells compared with parental cells. Moreover, overexpression of TCF12 sensitized DDP‐resistant cells to DDP by promoting ferroptosis. Intriguingly, silencing TCF12 reversed the promotion effect of the ferroptosis activator RSL3 on ferroptosis and DDP sensitivity, and overexpressing TCF12 antagonized the effect of the ferroptosis inhibitor liproxstatin‐1 on ferroptosis and DDP resistance. Mechanically, TCF12 promoted ubiquitination of SLC7A11 and decreased SLC7A11 protein stability through transcriptional repression of OTUB1, thereby facilitating ferroptosis. Consistently, SLC7A11 overexpression neutralized the promotion effect of TCF12 on ferroptosis and DDP sensitivity. Additionally, upregulation of TCF12 hindered the growth of mouse OSCC xenografts and enhanced the DDP sensitivity of xenografts by inducing ferroptosis. In conclusion, TCF12 enhanced DDP sensitivity in OSCC cells by promoting ferroptosis, which was achieved through modulating SLC7A11 expression via transcriptional regulation of OTUB1. [ABSTRACT FROM AUTHOR]

The study was to establish a fast and accurate ultra performance liquid chromatography-tandem mass spectrometry method for determination of DON in feed. The samples were extracted with 10 mL water, then oscillation and exocentric, purified by fungus toxin purification column, filtered by 0.22 μm membrane, determined by UPLC-MS/ MS. When DON supplementation levels were 0.01, 0.05 and 0.10 μg/kg, the recoveries were 83.92%~94.65% (concentrate feed), and RSD were 6.35%~7.09%. The DON had good linear relationships in their linear ranges, the linear coefficient was greater than 0.999 0, the limit of detection was 0.01 mg/kg. DON was detected in 194 of 216 batches of feed samples, and the detection rate was 89.8%. The study indicates that the method is easy to operate, low cost, high accuracy and can meet the requirements of large-scale detection of feed samples. [ABSTRACT FROM AUTHOR]